Skip to main content
Download PDF

Comprehensive analysis of the succinylome in Vero cells infected with peste des petits ruminants virus Nigeria 75/1 vaccine strain

BMC Veterinary Research volume 21, Article number: 45 (2025) Cite this article

Abstract

Background

Peste des petits ruminants virus (PPRV) is currently the only member of the Morbillivirus caprinae species within the genus Morbillivirus of the family Paramyoxviridae. PPRV causes a highly contagious disease in small ruminants, especially goats and sheep. Succinylation is a newly identified and conserved modification and plays an important role in host cell response to pathogen infection. However, the extent and function of succinylation in Vero cells during PPRV infection remains unknown.

Results

In this study, a global profile of the succinylome in Vero cells infected with PPRV Nigeria 75/1 vaccine strain (PPRVvac) was performed by dimethylation labeling-based quantitative proteomics analysis. A total of 2633 succinylation sites derived from 823 proteins were quantified. The comparative analysis of differentially succinylated sites revealed that 228 down-regulated succinylation sites on 139 proteins and 44 up-regulated succinylation sites on 38 proteins were significantly modified in response to PPRVvac infection, seven succinylation motifs were identified. GO classification indicated that the differentially succinylated proteins (DSuPs) mainly participated in cellular respiration, biosynthetic process and transmembrane transporter activity. KEGG pathway analysis indicated that DSuPs were related to protein processing in the endoplasmic reticulum. Protein-protein interaction networks of the identified proteins provided further evidence that various ATP synthase subunits and carbon metabolism were modulated by succinylation, while the overlapped proteins between succinylation and acetylation are involved in glyoxylate and dicarboxylate metabolism.

Conclusions

The findings of the present study provide the first report of the succinylome in Vero cells infected with PPRVvac and provided a foundation for investigating the role of succinylation alone and its overlap with acetylation in response to PPRVvac.

Peer Review reports

Background

Peste des petits ruminants virus (PPRV) is the etiological agent of peste des petits ruminants (PPR), which is a highly contagious transboundary disease and included in the World Organization of Animal Health (WOAH) list of notifiable terrestrial animal diseases. PPRV belongs to the Morbillivirus genus of the subfamily Orthoparamyxovirinae in the family Paramyoxviridae, alongside measles virus (MeV), rinderpest virus (RPV), canine distemper virus (CDV), phocine distemper virus (PDV), cetacean morbillivirus (CeMV) and feline morbillivirus (FeMV) [1,2,3]. PPRV has a negative, non-segmented single-stranded RNA genome encoding six structural proteins (N-P-M-F-H-L) and two non-structural proteins (V and C protein). PPRV was first reported in the Ivory Coast in 1942 and is now present in over 70 countries across Asia, Africa, Middle East and Europe, inhibiting trade and causing significant economic losses.

Host proteins are highly contested targets in the ongoing “arms race” between viruses and the host. Viruses “hijack” host cellular functions to facilitate their replication and inhibit host antiviral defenses. On the contrary, the host actively mobilizes the immune antiviral response to resist viral invasion, or inhibit the replication of viral particles, or eliminate virus particles. Today, it is well known that protein post-translational modifications (PTMs) significantly affect the diverse function of proteins through the modulation of biological processes, protein activity, cellular location and protein-protein interaction (PPI) by transferring modified groups to one or more amino acid residues. In recent years, protein PTMs have become a hot topic in viral infection.

To date, over 450 protein modifications including more than 200 PTMs have been identified [4]. They are dynamic and reversible protein processing events that play key roles in disease pathogenesis [5,6,7,8]. Some PTMs including phosphorylation, methylation, acetylation and succinylation have been shown to potently regulate innate immunity and inflammation in response to virus infections [9, 10]. Of the 20 amino acid residues, lysine is a frequent target of covalent modifications because it can accept different types of chemical groups. Succinylation is a newly discovered and meagerly studied modification that is evolutionarily conserved from bacteria to mammals. It can transfer a larger structural moiety (a succinyl group, -CO-CH2-CH2-CO2H) than that in acetylation, and changes the charge on the modified residues from + 1 to − 1 causing a two-unit charge shift in the modified residues. Consequently, the structure and function of the succinylated proteins might have greater changes, which in turn results in substantial changes in the chemical properties of the target proteins indicating that succinylation can potentially regulate the protein structure and function associated with diverse cellular processes such as metabolism and translation [11,12,13].

Lysine succinylation was first identified as a new PTM in 2011 [7]. So far, succinylation is widespread in diverse species ranging from prokaryotes and eukaryotes, from plant to animal [4, 14,15,16,17]. It causes unique functional consequences and affects chromatin structure, conformational stability and gene expression [13, 18]. In recent years, more and more attention has been paid to the high coincidence of lysine succinylation modification with acetylation modification [17, 19,20,21,22,23,24]. The majority of succinylation sites in bacteria, yeast and mammal cell were acetylated at the same position, which suggest that these two modifications may affect the performance of proteins together. The coincidence sites of lysine succinylation were also mostly located in polar, acid or alkaline amino acid regions, and were exposed to protein surface [13, 17, 25]. Moreover, succinylation occurs at a low level and as such, many succinylation sites remain unidentified.

Despite the advanced findings at a quick pace with the developments in mass spectrometry (MS) technologies and peptide enrichment methods, global succinylome and the relationship between succinylation and acetylation remain an understudied aspect of PPRV infection. Therefore, systematical analyses of host cellular protein succinylation might provide helpful insights into PPRV pathogenesis and replication that could lead to the discovery of antiviral targets.

In the present study, we aimed to investigate the succinylome of African green monkey kidney (Vero) cells (ATCC; CCL-81) with or without PPRV Nigeria 75/1 vaccine strain (PPRVvac) infection, by combining dimethylation labeling and LC-MS/MS analysis. We successfully quantified 2633 succinylation sites on 823 proteins with diverse molecular functions, biological processes and subcellular localizations. 272 succinylation sites on 177 proteins were significant modified in response to PPRVvac, seven lysine succinylation motifs were identified. PPI networks further indicated that various ATP synthase subunits and carbon metabolism were modulated by succinylation, the overlapped proteins between succinylation and acetylation were involved in glyoxylate and dicarboxylate metabolism. Overall, the results provided the first extensive dataset of succinylation in Vero cells infected by PPRVvac, and a novel view on investigating the mechanism of PPRV infection.

Results

Global detection of lysine succinylated sites on PPRVvac-infected Vero cellular proteins

To explore and identify the host proteins with succinylated sites involved in PPRVvac replication, we detected the abundance of succinylated proteins in PPRVvac-infected Vero cells by western blotting and selected 24 hpi as the time point for quantitative proteomic analysis (Fig. S1). The near-zero distribution of mass error and that the errors were predominantly < 0.02 Da (Fig. S2). Most of the enriched succinylated peptide lengths were in the range of 7–21 segments (Fig. 1A), which was consistent with trypsin cutting at lysine residue sites.

Fig. 1
figure 1

The identification of succinylation proteins and sites. A peptide length distribution of succinylation profile; B Summary of the succinylated proteins and sites identified

Full size image

A total of 2840 succinylated sites across 875 proteins were identified, of which 2633 sites on 823 proteins were quantified (Table S1). The number of succinylated sites per protein varies from 1 to 33 depending on the protein. Among these proteins, about 383 (43.77%) included a single succinylation site, 176 (20.11%) included two succinylation sites, 86 (9.83%) included three succinylation sites, 62 (7.09%) included four succinylation sites, and 168 (19.20%) included five or more than five succinylation sites (Fig. 1B). 28 succinylation sites were found on histone proteins, including 1 site in H1C, 2 sites in H2A, 16 sites in H2B, 4 sites in H3 and 5 sites in H4.

Based on a threshold of 1.2 fold changes and t test P < 0.05 as standards, among the 2633 quantifiable sites, 272 succinylated sites on 177 proteins can response to PPRVvac infection (Table S2). 44 succinylated sites on 38 proteins were quantified as significantly upregulated, and 228 succinylated sites on 129 proteins were significantly downregulated. Most of these differentially succinylated proteins (DSuPs) were modified at a single succinylation site, including clathrin heavy chain (K557), CCT3 (K353), CCT5 (K223), ezrin (K57), plectin (K3390) and NLRX1 (K656). 58 DSuPs were modified at multiple sites, including heat-shock protein (HSP) A9 (7 sites), serine hydroxymethyltransferase 2 (SHMT2, 6 sites), HSPA5 (4 sites), HSP90AB1 (3 sites), HSPA8 (2 sites), isocitrate dehydrogenase 2 ( IDH2, 2 sites), Isocitrate dehydrogenase isocitrate dehydrogenase 3 beta subunit (IDH3B, 2 sites) and vimentin (2 sites).

Motifs of succinylated peptides

To examine the properties of the succinylated lysine in PPRVvac-infected Vero cells, the sequences from the − 10 to + 10 positions flanking the succinylated sites were investigated using the Motif-X with a significance threshold of P < 0.000001. The amino acids flanking the succinylated sites were matched to the whole size.

Of all succinylated peptides, seven significantly enriched lysine succinylation site motifs from 1310 modified sites were identified (Fig. 2A). These motifs were AKsc, Ksc******K, Ksc*******K, Ksc***K*****V, A****Ksc, Ksc******V and V*Ksc (Ksc: succinylated lysine; *: residue of a random amino acid). According to the position and other properties of the residues around the succinylated lysine, lysine (K) at + 7/+8 or alanine (A) at − 1 position was more readily succinylated, the frequency of K at + 4 and valine (V) at + 10 position was the lowest. The positively charged lysine (K) residue was enriched at the positions + 4 to + 8. The motif enrichment was illustrated in the form of a heat map (Fig. 2B).

Fig. 2
figure 2

Characterization of succinylated peptides. A Probability sequence motifs of succinylation sites consisting of 20 residues surrounding the targeted lysine residue using Motif-X. Seven significantly enriched succinylation site motifs were identified; B Heat map showing upstream (red) or downstream (green) of amino acid compositions around the succinylated lysine site (10 amino acids upstream and downstream of the succinylated lysine site)

Full size image

Functional classification of differentially succinylated proteins

DSuPs were classified by Gene Ontology (GO) functional classification analysis and COG/KOG categories (Table S3). In the biological processes, three major classes of succinyl-proteins were associated with metabolic, cellular and single-organism processes, accounting for 35%, 29% and 23% of the total DSuPs, respectively (Fig. 3A). Cell (41%), organelle (22%) and membrane (20%) were identified as favorable cellular components (Fig. 3B). The succinylated proteins were mostly related to binding catalytic activity (49%) and binding (37%) (Fig. 3C). Most of DSuPs were more abundant in mitochondria (54%), cytoplasm (25%), extracellular (6%) and nucleus (6%) (Fig. 3D). Moreover, 166 DSuPs were successfully annotated into four categories (Fig. 4). 78 DSuPs (47%) were involved in metabolism, and 41% played roles in energy production and conversion. 41 DSuPs (25%) were related to cellular processes and signaling, and 71% were involved in post-translational modification, protein turnover and chaperones.

Fig. 3
figure 3

Classification of the identified DSuPs. A Biological process; B Cellular component; C Molecular function; D Subcellular localization

Full size image
Fig. 4
figure 4

COG/KOG classification

Full size image

Enrichment of differentially succinylated proteins

To further explore the functions of the succinylated proteins during PPRVvac infection, GO, Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein domain enrichment were performed for all the identified DSuPs (Fig. 5, Table S4).

In the molecular function category, transmembrane transporter activity, unfolded protein binding, isomerase activity and oxidoreductase activity were found to be significantly enriched (Fig. 5A). The most enriched cellular component was the proton-transporting ATP synthase complex (Fig. 5B). The succinylated proteins were significantly enriched in the cellular metabolic process with specific enrichment in cellular respiration, glycosyl compound biosynthetic process and ribose phosphate biosynthetic process (Fig. 5C). 13 DSuPs were involved in protein processing in the endoplasmic reticulum (ER), and 9 of which were down-regulated. Furthermore, the protein domain analysis showed that a large proportion of DSuPs were associated with the ClpP/crotonase-like domain and ribosomal protein S5 (RPS5) domain 2-type fold (Fig. 5D).

Fig. 5
figure 5

Enrichment of DSuPs. A Molecular function; B Cellular component; C Biological process. D: Protein domain

Full size image

Protein–protein interaction network analysis

PPIs are critical for various biological processes. To better understand how succinylation regulates diverse metabolic processes and cellular functions in PPRVvac-infected Vero cells, PPI analysis was performed by searching the STRING database and PPI networks were visualized using Cytoscape software.

PPI network included 108 succinylated proteins as nodes, connected by 804 identified interactions (Fig. 6, Table S5). Two highly connected subnetworks, carbon metabolism and protein processing in the ER of DSuPs were enriched. In the first subnetwork, 16 succinylated proteins with 133 Ksc sites and 215 direct physical interactions participated in carbon metabolism, suggesting that they could play key roles in energy transformation. Among the 16 succinylated proteins, four (γ, β, O and δ) subunits of ATP synthases were identified as DSuPs in PPRVvac-infected cells. The second subnetwork comprised 6 succinylated proteins with 142 Ksc sites and 39 direct physical interactions and were related to protein disulfide-isomerase (PDI) and HSP. In these two subnetworks, protein–protein interaction networks including 3 up-regulated proteins and 19 down-regulated proteins were significantly enriched. Most of the proteins in the PPI network contain more than two succinylated sites.

Fig. 6
figure 6

PPI networks of DSuPs. Each node represented a protein, and each line referred to an interaction. Node colors represented fold change, and circle size represents the numbers of DSuPs, and red indicates upregulated and blue indicates downregulated DSuPs

Full size image

To determine if the succinylation and acetylation occur at the same lysine site, we compared the succinylation data to the acetylation data (ProteomeXchange, PXD025081) of modification sites and proteins. The results showed that 34 DSuPs overlapped with differentially acetylated proteins (DAcPs), one highly connected subnetwork, glyoxylate and dicarboxylate metabolism were enriched (Fig. 7). 18 succinylated sites of 13 DSuPs were also acetylated on the same lysine. Of these 13 DSuPs, 8 proteins had only one modification site, while 5 proteins had two modification sites.

Fig. 7
figure 7

The crosstalk between the DSuPs and DAcPs in PPRVvac-infected vero cell

Full size image

Discussion

Host infected with pathogen usually undergoes a series of physiological and biochemical changes at the cellular and molecular levels. It can lead to major changes during transcriptional, post-transcriptional, translational and post-translational levels. As a conserved PTM, lysine succinylation is widespread in diverse species [4, 14,15,16,17], and hundreds of protein succinylation sites are present and being actively investigated. Succinylation has a negatively charged carboxylate group that results in more distinct chemical and structural changes in lysine residues. Succinylation plays a central role in regulating multiple biological processes, especially for metabolism [26]. The changes in metabolism alter succinylation, then succinylation provides feedback on metabolism and crosstalk with other proteins that are important to pathology. Succinylation was initially discovered in mitochondrial proteins involved in the TCA cycle, fatty acid metabolism, and amino acid degradation [27]. It implies that succinylation may help to integrate the responses to the diverse metabolic challenges. Nonetheless, Succinylation modulates the activities of enzymes and pathways by modifying catalysis or cofactor binding sites. Succinylation is also associated with pathophysiology and diseases as well as physiological processes [28].

All PPR vaccines currently in use are live attenuated strains and belong to either lineage II or lineage IV. Live cell culture attenuated PPR vaccine seems to have the same characteristic as the wild type strains and appears able to protect against any naturally occurring strain of the virus. Up to date, although comparative transcriptome and proteomic profiling were analysed in bone marrow-derived dendritic cells (BMDCs) or peripheral blood mononuclear cells (PBMCs) stimulated with PPRV (vaccine strains Sungri/96 and Nigeria 75/1) [29,30,31], little is known about lysine succinylation in bio-engineering cells infected with PPRVvac. In the present study, for the first time, we elucidated the effects of PPRVvac infection on the protein succinylation profiles of Vero cells. This systematic analysis will provide an important baseline for functional studies on the response of the succinylated proteins to PPRVvac, and will be helpful to elucidate the cellular events occurred in Vero cells following field virus stimulation.

Overall, we identified 2840 succinylation sites in 875 cellular proteins, and 2633 modification sites in 823 proteins regulated in PPRVvac-infected Vero cells. 177 proteins with 272 succinylation sites were differentially succinylated in response to PPRVvac. Of these DSuPs, substrates (SHMT2, IDH2, IDH3B) of sirtuin 5 (Sirt5), HSPs, ATP synthase subunits, PDIA4 and vimentin were modified at multiple succinylation sites. Sirt5 has weak deacetylase activity but can catalyze the removal of three acidic lysine modifications, malonylation, succinylation and glutarylation [32], and is the only desuccinylase located in the cytoplasm [27, 33]. By desuccinylating substrates, Sirt5 regulates a variety of biological processes, such as active oxygen defense, fatty acid metabolism, cell apoptosis and autophagy [34,35,36]. Notably, PPRVvac induces and exploits autophagy for replication via distinct signaling pathways [37,38,39]. All succinylated sites of SHMT2, IDH2, IDH3B were significantly down-regulated in this study. Hereby, we hypothesized that desuccinylated SHMT2, IDH2, IDH3B may be promote PPRVvac replication by inducing autophagy. HSP70 isoforms such as HSPA5, HSPA8 and HSPA9 are required at all steps of the virus life cycle [40,41,42]. Vimentin plays important roles in viral infection by interacting with viral proteins [43,44,45,46,47,48]. Although NLRX1 only has one succinylated site in the study, it is virtually worth researching in immune system function of virus infection. NLRX1 can negatively regulate type-I interferon, attenuate pro-inflammatory NF-kB signaling and modulate autophagy, cell death, and proliferation. NLRX1 can interacts with virus protein to enhance or restrict viral replication [49, 50]. However, the DSuPs with differentially expression levels were rare in this study, which suggested that the key roles of the succinylated proteins in PPRVvac infection are determined by their succinylation levels. Moreover, the numbers of succinylated sites and proteins were less than that of acetylation, which implied that acetylation may be more important than succinylation in PPRVvac infection.

Succinylation is a charged PTM that modifies a charged residue, making more hydrophobic regions unlikely targets for modification. The different desuccinylases have a varied sequence of bias towards specific lysine-succinylation sites. The motifs found in this study line up with the previous literatures [51, 52]. The positively charged K amino acid at the positions + 4 to + 8 was enriched and succinylated. A and V amino acids were also preference residues around succinylation sites. These results showed that K residue with aliphatic amino acids is the preferred substrate for succinyl group donor, such as succinyl-CoA.

The results of GO functional classification indicated that the identified DSuPs are involved in catalytic activity and binding. Proteins related to proton-transporting ATP synthase complex were highly enriched, which consistent with the molecular function category and protein domain enrichment results. ClpP/crotonase plays an important role in protein quality and homeostasis in the cell by functioning mainly in the disaggregation, unfolding and degradation of native as well as misfolded proteins [53]. RPS5 plays an essential role in protein translation and is involved in cell differentiation and apoptosis [54, 55]. RPS5 combines with the internal ribosome entry site (IRES) of hepatitis virus to provide the initial stages of translation initiation on the viral RNA [56]. However, the molecular mechanism of RPS5 is still unclear. ER is critical for protein synthesis and maturation and relies on many molecular chaperones that assist in protein folding and assembly. Virus infection can alter ER and activate the unfolded-protein response to facilitate viral replication [57, 58]. The KEGG enrichment analysis showed that 13 of DSuPs might play a major role in protein processing in the ER of PPRVvac-infected Vero cells. Therefore, the relationship between PPRVvac infection and DSuPs requires further investigations.

PPIs are critical for various biological processes. The present study provided the first global PPI network of DSuPs in Vero cells induced by PPRVvac. The subnetworks of carbon metabolism were enriched. ATP synthases are membrane protein complexes closely related to respiration in mitochondria. They are molecular motors and can synthesize ATP by continuous rotation, and maintain the energy needed for metabolism. In the second subnetwork, some members of PDI and HSP were enriched. PDI can catalyze the formation, breakage and rearrangement of disulfide bonds, and promote protein folding, which is essential to stabilize the three-dimensional structure of proteins. The imbalance of PDI expression or enzyme activity is closely related to a series of diseases. HSPs play various physiological functions by binding to protein molecules. HSPs can help amino acid chains fold into the correct three-dimensional structure, clean up the mis-folded proteins, escort proteins to find target molecules. HSPs also participate in innate immune response. These results suggest that the succinylation of these proteins play an important role in protein anabolism of PPRVvac-infected Vero cells. Moreover, 18 succinylation sites on 34 DSuPs overlapped with acetylation sites. Succinylated sites at the 129 th of vimentin, 81 st and 113 th of HSPA5, 56 th and 601 st of HSPA8, 394 th and 600 th of HSPA9, 112 nd and 496 th of HSPD1 overlapped with acetylation sites. Minimal overlap of succinylation and acetylation sites indicates differential regulation of succinylation and acetylation [27]. However, others reported that succinylation occurs at a low level and extensively overlaps with acetylation in prokaryotes and eukaryotes [17, 59]. A comprehensive study is required to determine whether the relationship between succinylation and acetylation occurring on the same lysine in PPRVvac infection is independent or competitive, or synergistic. Thus, these results provided a promising starting point for further characterizing the regulatory roles of succinylation alone and the crosstalk between lysine succinylation and acetylation in response to PPRVvac. This will be helpful to enhance understanding of the interplay between PTMs and PPRV and provide valuable clues for identification of the potential molecular targets of PPRV among cellular proteins.

Conclusion

In this study, we identified 272 succinylated sites of 177 DSuPs response to PPRVvac infection. These succinylated proteins primarily participated in cellular respiration and biosynthetic process, providing insights into metabolism and succinylation after PPRVvac infection. PPI networks further indicated that various ATP synthase subunits and carbon metabolism were modulated by succinylation, the overlapped proteins between succinylation and acetylation are involved in glyoxylate and dicarboxylate metabolism. The overlap between DSuPs and DAcPs implies the possibility of crosstalk between these two PTMs in response to PPRVvac infection. Further research is needed to evaluate the regulatory role of DSuPs and DAcPs involved in the host cell response to PPRVvac.

Methods

Cell, virus and infection

African green monkey kidney (Vero) cells (ATCC; CCL-81) were maintained in authors’ laboratory, and cultured in DMEM medium (Sigma Aldrich, St Louis, MO, USA) supplemented with 10% fetal bovine serum, 100 IU/ml penicillin and 100 µg/ml streptomycin and incubated at 37 °C in 5% CO2 incubator. The PPRVvac was cultured in our laboratory, which was propagated by Vero cells [60]. Vero cells seeded in 6-well cell culture plates were infected with PPRVvac at a MOI of 0.1 or mock-infected with phosphate-buffered saline (PBS, 0.01 M, pH7.4) and incubated at 37 °C for 1 h. The MOI was determined based on the viral titers of Vero cell line. After infection, the virus inoculum was removed, and the fresh medium was then added to wells and incubated. The infected cells were harvested at 24, 48, 72 h post infection (hpi) and analysed using western blotting (20 µg/sample) and a pan anti-succinyllysine antibody (diluted at 1:1000; PTM-419, PTM Biolabs, Hangzhou, China), and non-inoculated cells served as control group. Three independent biological replicates (three parallel experiments) were performed. The flowchart of the present study was shown in Fig. 8.

Fig. 8
figure 8

The flowchart of virus infection, proteomic analysis and global mapping of succinylation in PPRVvac-infected Vero cells

Full size image

Protein extraction

The harvested cell samples were washed twice with cold phosphate-buffered saline (PBS). Then, each sample was sonicated on ice in lysis buffer (8 M urea, 1% protease inhibitor cocktail, 3 µM trichostatin A (TSA), 50 mM nicotinamide (NAM) and 2 mM EDTA). The resulting supernatants were centrifuged with 12,000 rpm for 10 min at 4 °C to remove the cellular debris. The protein concentration was determined with BCA kit according to the manufacturer’s instructions.

Trypsin digestion and dimethylation labeling

The protein solution was reduced with 5 mM dithiothreitol for 30 min at 56 °C and subsequently alkylated with 11 mM iodoacetamide for 15 min at room temperature in darkness. For tryptic digestion, the protein samples were diluted to urea concentration less than 2 M. Finally, trypsin was added at 1:50 trypsin-to-protein mass ratio for the first digestion overnight and at 1:100 trypsin-to-protein mass ratio for a second 4 h-digestion at 37 °C. Then, the peptide was desalted using Strata X C18 SPE column (Phenomenex) and vacuum-dried. Peptide samples were resuspended in 0.1 M TEAB and and formaldehyde/deuterated formaldehyde were further treated with sodium cyanoborohydride, submitted to incubation for 2 h at room temperature, desalting, and drying.

Enrichment of succinylated peptides

Lysine-succinylated peptides were enriched with an agarose-conjugated pan anti-succinyllysine antibody (PTM Biolabs, Hangzhou, China). In brief, dried tryptic peptides dissolved in NETN buffer (100 mM NaCl, 1 mM EDTA, 50 mM Tris-HCl, 0.5% NP-40, pH 8.0) were incubated with pre-washed pan anti-succinyllysine (PTM-402, PTM Biolabs, Hangzhou, China) conjugated agarose beads at 4 °C overnight with gentle oscillation. The beads were washed four times with NETN buffer and twice with ice-cold ddH2O. The bound peptides were eluted three times from the beads with 0.1% trifluoroacetic acid (TFA; Sigma-Aldrich, St. Louis, MO, USA). The eluted fractions were pooled together and vacuum-dried. The resulting peptides were desalted using C18 ZipTips (Merck Millipore, Billerica, MA, USA) according to the manufacturer’s instructions and dried by vacuum centrifugation.

LC-MS/MS analysis

The enriched peptides were dissolved in solvent A (0.1% Formic Acid (FA) in 2% acetonitrile (ACN)), and directly loaded onto a home-made reversed-phase analytical column (1.9 μm beads, 120 A pore, 15-cm length, 75 μm i.d.). The gradient was comprised of a four-step linear: from 9 to 25% solvent B (0.1% FA in 90% ACN) for 24 min, from 25 to 40% for 10 min, from 40 to 80% for 3 min, and maintained at 80% for the last 3 min on an EASY-nLC 1000 UPLC (Ultra Performance Liquid Chromatography) system at a constant flow rate of 700 nL/min.

The peptides eluting from the analytical column were ionized and subjected to tandem mass spectrometry (MS/MS) in a Q Exactive™ Plus (Thermo Scientific) coupled online to the UPLC using a NanoSpray Ionization (NSI) source. The applied electrospray voltage was 2.0 kV. Intact peptides were detected at a resolution of 70,000 with a scan range of 350–1800 m/z for full MS scans using Orbitrap. Peptides were then selected for MS/MS using an NCE setting of 28 and ion fragments were detected at a resolution of 17,500. Data-dependent acquisition (DDA) that alternated between one MS scan followed by 20 MS/MS scans was applied for the top 20 precursor ions, with 15 s dynamic exclusion. Automatic gain control (AGC) was used to prevent overfilling of the ion trap and was set at 5E4, with a fixed first mass of 100 m/z. The MS proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD025025.

Database searches

The protein succinylation sites identification and quantification were processed using MaxQuant search engine (v.1.5.2.8) against the Chlorocebus sabaeus database (19,228 sequences), concatenated with a reverse database and common contaminants. Trypsin/P was specified as a cleavage enzyme allowing up to 4 missing cleavages. The mass tolerance for the precursor ions was set to 20 ppm in the first analysis and 5 ppm in the full search, and the fragment mass tolerance was set to 0.02 Da. Carbamido-methylation of cysteine (Cys) was specified as a fixed modification, and oxidation of methionine, succinylation on the protein N-terminal and succinylation on lysine were specified as variable modifications. The false discovery rate (FDR) thresholds and the minimum score for the modified peptide were set to < 1% and > 40, respectively. The minimum peptide length was set to seven. All other parameters in MaxQuant were set to default values.

Bioinformatics analysis

GO annotation was derived from the UniProt-GOA database (http://www.ebi.ac.uk/GOA). The proteins were classified into three categories: biological processes, cellular components and molecular functions. Protein domain annotation was performed using InterProScan (http://www.ebi.ac.uk/InterProScan/) based on the protein sequence alignment method, and the InterPro domain database (http://www.ebi.ac.uk/interpro/). The KEGG database (http://www.genome.jp/kegg/) was used to annotate and map the pathways. GO, protein domain and KEGG pathway enrichment analysis were performed using DAVID bioinformatics resources 6.8. Wolfpsort (https://wolfpsort.hgc.jp/), a subcellular localization predication soft was used to predict subcellular localization. Amino acid sequence motifs (within ± 10 residues of the succinylated sites) were analysed using Motif-X. Motif-based clustering analyses were also performed, and cluster membership was visualized using a heat map. Functional interaction network analysis was performed using the STRING database (v.11.0), with a threshold of 0.7 (high confidence) and visualized using Cytoscape 3.7.1.

Data availability

All data generated or analysed during this study are included in this published article. And The LC–MS/MS proteomics data have been deposited in the ProteomeXchange Consortium with the dataset identifier PXD025025.

Abbreviations

PPRV:

Peste des petits ruminants virus

PPRVvac:

PPRV Nigeria 75/1 vaccine strain

PPR:

Peste des petits ruminants

DSuPs:

Differentially succinylated proteins

PTMs:

Post-translational modifications

PDI:

Protein disulfide-isomerase

HSP:

Heat-shock protein

PPI:

Protein-protein interaction

DAcPs:

Differentially acetylated proteins

References

  1. Liu F, Li J, Li L, Liu Y, Wu X, Wang Z. Peste Des petits ruminants in China since its first outbreak in 2007: a 10-year review. Transbound Emerg Dis. 2018;65(3):638–48.

    Article  CAS  PubMed  Google Scholar 

  2. Woo PC, Lau SK, Wong BH, Fan RY, Wong AY, Zhang AJ, et al. Feline morbillivirus, a previously undescribed paramyxovirus associated with tubulointerstitial nephritis in domestic cats. Proc Natl Acad Sci U S A. 2012;109(14):5435–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Kennedy S. Morbillivirus infections in aquatic mammals. J Comp Pathol. 1998;119(3):201–25.

    Article  CAS  PubMed  Google Scholar 

  4. Zhen S, Deng X, Wang J, Zhu G, Cao H, Yuan L, et al. First Comprehensive Proteome Analyses of Lysine Acetylation and Succinylation in Seedling leaves of Brachypodium distachyon L. Sci Rep. 2016;6:31576.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Yang F. Post-translational modification control of HBV Biological processes. Front Microbiol. 2018;9:2661.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Chen L, Keppler OT, Scholz C. Post-translational modification-based regulation of HIV replication. Front Microbiol. 2018;9:2131.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Zhang Z, Tan M, Xie Z, Dai L, Chen Y, Zhao Y. Identification of lysine succinylation as a new post-translational modification. Nat Chem Biol. 2011;7(1):58–63.

    Article  CAS  PubMed  Google Scholar 

  8. Choudhary C, Kumar C, Gnad F, Nielsen ML, Rehman M, Walther TC, et al. Lysine acetylation targets protein complexes and co-regulates major cellular functions. Science. 2009;325(5942):834–40.

    Article  CAS  PubMed  Google Scholar 

  9. Zhou Y, He C, Wang L, Ge B. Post-translational regulation of antiviral innate signaling. Eur J Immunol. 2017;47(9):1414–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Liu J, Qian C, Cao X. Post-Translational Modif Control Innate Immun Immun. 2016;45(1):15–30.

    Google Scholar 

  11. Yao Z, Guo Z, Wang Y, Li W, Fu Y, Lin Y, et al. Integrated Succinylome and Metabolome Profiling reveals crucial role of S-Ribosylhomocysteine lyase in Quorum sensing and metabolism of Aeromonas hydrophila. Mol Cell Proteom. 2019;18(2):200–15.

    Article  CAS  Google Scholar 

  12. Zhou H, Finkemeier I, Guan W, Tossounian MA, Wei B, Young D, et al. Oxidative stress-triggered interactions between the succinyl- and acetyl-proteomes of rice leaves. Plant Cell Environ. 2018;41(5):1139–53.

    Article  CAS  PubMed  Google Scholar 

  13. Yang M, Wang Y, Chen Y, Cheng Z, Gu J, Deng J, et al. Succinylome analysis reveals the involvement of lysine succinylation in metabolism in pathogenic Mycobacterium tuberculosis. Mol Cell Proteom. 2015;14(4):796–811.

    Article  CAS  Google Scholar 

  14. Xie L, Li J, Deng W, Yu Z, Fang W, Chen M, et al. Proteomic analysis of lysine succinylation of the human pathogen Histoplasma Capsulatum. J Proteom. 2017;154:109–17.

    Article  CAS  Google Scholar 

  15. Li X, Hu X, Wan Y, Xie G, Chen D, Cheng Z, et al. Systematic identification of the lysine succinylation in the protozoan parasite Toxoplasma Gondii. J Proteome Res. 2014;13(12):6087–95.

    Article  CAS  PubMed  Google Scholar 

  16. Colak G, Xie Z, Zhu AY, Dai L, Lu Z, Zhang Y, et al. Identification of lysine succinylation substrates and the succinylation regulatory enzyme CobB in Escherichia coli. Mol Cell Proteom. 2013;12(12):3509–20.

    Article  CAS  Google Scholar 

  17. Weinert BT, Scholz C, Wagner SA, Iesmantavicius V, Su D, Daniel JA, et al. Lysine succinylation is a frequently occurring modification in prokaryotes and eukaryotes and extensively overlaps with acetylation. Cell Rep. 2013;4(4):842–51.

    Article  CAS  PubMed  Google Scholar 

  18. Xie Z, Dai J, Dai L, Tan M, Cheng Z, Wu Y, et al. Lysine succinylation and lysine malonylation in histones. Mol Cell Proteom. 2012;11(5):100–7.

    Article  CAS  Google Scholar 

  19. Mizuno Y, Nagano-Shoji M, Kubo S, Kawamura Y, Yoshida A, Kawasaki H, et al. Altered acetylation and succinylation profiles in Corynebacterium glutamicum in response to conditions inducing glutamate overproduction. Microbiologyopen. 2016;5(1):152–73.

    Article  CAS  PubMed  Google Scholar 

  20. Zeng J, Wu L, Chen Q, Wang L, Qiu W, Zheng X et al. Comprehensive profiling of protein lysine acetylation and its overlap with lysine succinylation in the Porphyromonas gingivalis fimbriated strain ATCC 33277. Mol Oral Microbiol. 2020.

  21. Kosono S, Tamura M, Suzuki S, Kawamura Y, Yoshida A, Nishiyama M, et al. Changes in the Acetylome and Succinylome of Bacillus subtilis in response to Carbon source. PLoS ONE. 2015;10(6):e0131169.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Pan J, Chen R, Li C, Li W, Ye Z. Global Analysis of Protein Lysine Succinylation Profiles and their overlap with lysine acetylation in the Marine Bacterium Vibrio parahemolyticus. J Proteome Res. 2015;14(10):4309–18.

    Article  CAS  PubMed  Google Scholar 

  23. Rardin MJ, He W, Nishida Y, Newman JC, Carrico C, Danielson SR, et al. SIRT5 regulates the mitochondrial lysine succinylome and metabolic networks. Cell Metab. 2013;18(6):920–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Cao Y, Fan G, Wang Z, Gu Z. Phytoplasma-induced changes in the Acetylome and Succinylome of Paulownia tomentosa provide evidence for involvement of Acetylated Proteins in witches’ broom disease. Mol Cell Proteom. 2019;18(6):1210–26.

    Article  CAS  Google Scholar 

  25. Rosen R, Becher D, Buttner K, Biran D, Hecker M, Ron EZ. Probing the active site of homoserine trans-succinylase. FEBS Lett. 2004;577(3):386–92.

    Article  CAS  PubMed  Google Scholar 

  26. Peng C, Lu Z, Xie Z, Cheng Z, Chen Y, Tan M, et al. The first identification of lysine malonylation substrates and its regulatory enzyme. Mol Cell Proteom. 2011;10(12):M111012658.

    Article  Google Scholar 

  27. Park J, Chen Y, Tishkoff DX, Peng C, Tan M, Dai L, et al. SIRT5-mediated lysine desuccinylation impacts diverse metabolic pathways. Mol Cell. 2013;50(6):919–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Hirschey MD, Zhao Y. Metabolic regulation by Lysine Malonylation, Succinylation, and glutarylation. Mol Cell Proteom. 2015;14(9):2308–15.

    Article  CAS  Google Scholar 

  29. Manjunath S, Mishra BP, Mishra B, Sahoo AP, Tiwari AK, Rajak KK, et al. Comparative and temporal transcriptome analysis of peste des petits ruminants virus infected goat peripheral blood mononuclear cells. Virus Res. 2017;229:28–40.

    Article  CAS  PubMed  Google Scholar 

  30. Li L, Wu J, Cao X, Zhou J, Yin S, Yang S, et al. Proteomic analysis of murine bone marrow derived dendritic cells in response to peste des petits ruminants virus. Res Vet Sci. 2019;125:195–204.

    Article  CAS  PubMed  Google Scholar 

  31. Li L, Wu J, Liu D, Du G, Liu Y, Shang Y et al. Transcriptional profiles of murine bone marrow-derived dendritic cells in response to Peste Des Petits ruminants Virus. Vet Sci. 2019; 6(4).

  32. Du J, Zhou Y, Su X, Yu JJ, Khan S, Jiang H, et al. Sirt5 is a NAD-dependent protein lysine demalonylase and desuccinylase. Science. 2011;334(6057):806–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Anderson KA, Green MF, Huynh FK, Wagner GR, Hirschey MD, SnapShot. Mammalian Sirtuins Cell. 2014;159(4):956–e956951.

    CAS  PubMed  Google Scholar 

  34. Zhang Y, Li T, Cai X, Long D, Wang X, Liu C, et al. Sirt5-mediated desuccinylation of OPTN protects retinal ganglion cells from autophagic flux blockade in diabetic retinopathy. Cell Death Discov. 2022;8(1):63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Shi L, Yan H, An S, Shen M, Jia W, Zhang R, et al. SIRT5-mediated deacetylation of LDHB promotes autophagy and tumorigenesis in colorectal cancer. Mol Oncol. 2019;13(2):358–75.

    Article  CAS  PubMed  Google Scholar 

  36. Garva R, Thepmalee C, Yasamut U, Sudsaward S, Guazzelli A, Rajendran R, et al. Sirtuin Family members selectively regulate autophagy in Osteosarcoma and Mesothelioma Cells in response to Cellular stress. Front Oncol. 2019;9:949.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Zhang R, Lin H, You Q, Zhang Z, Bai L, Chen F, et al. Peste Des Petits ruminants Virus Upregulates STING to activate ATF6-Mediated autophagy. J Virol. 2022;96(20):e0137522.

    Article  PubMed  Google Scholar 

  38. Yang B, Xue Q, Guo J, Wang X, Zhang Y, Guo K, et al. Autophagy induction by the pathogen receptor NECTIN4 and sustained autophagy contribute to peste des petits ruminants virus infectivity. Autophagy. 2020;16(5):842–61.

    Article  CAS  PubMed  Google Scholar 

  39. Zhang Y, Wu S, Lv J, Feng C, Deng J, Wang C, et al. Peste Des petits ruminants virus exploits cellular autophagy machinery for replication. Virology. 2013;437(1):28–38.

    Article  CAS  PubMed  Google Scholar 

  40. Taguwa S, Yeh MT, Rainbolt TK, Nayak A, Shao H, Gestwicki JE, et al. Zika Virus dependence on host Hsp70 provides a protective strategy against infection and disease. Cell Rep. 2019;26(4):906–e920903.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Su YS, Hsieh PY, Li JS, Pao YH, Chen CJ, Hwang LH. The heat shock protein 70 family of chaperones regulates all phases of the Enterovirus A71 Life Cycle. Front Microbiol. 2020;11:1656.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Iyer K, Chand K, Mitra A, Trivedi J, Mitra D. Diversity in heat shock protein families: functional implications in virus infection with a comprehensive insight of their role in the HIV-1 life cycle. Cell Stress Chaperones. 2021;26(5):743–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Kim JK, Fahad AM, Shanmukhappa K, Kapil S. Defining the cellular target(s) of porcine reproductive and respiratory syndrome virus blocking monoclonal antibody 7G10. J Virol. 2006;80(2):689–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Das S, Ravi V, Desai A. Japanese encephalitis virus interacts with vimentin to facilitate its entry into porcine kidney cell line. Virus Res. 2011;160(1–2):404–8.

    Article  CAS  PubMed  Google Scholar 

  45. Du N, Cong H, Tian H, Zhang H, Zhang W, Song L, et al. Cell surface vimentin is an attachment receptor for enterovirus 71. J Virol. 2014;88(10):5816–33.

    Article  PubMed  PubMed Central  Google Scholar 

  46. Yu YT, Chien SC, Chen IY, Lai CT, Tsay YG, Chang SC, et al. Surface vimentin is critical for the cell entry of SARS-CoV. J Biomed Sci. 2016;23:14.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Risco C, Rodriguez JR, Lopez-Iglesias C, Carrascosa JL, Esteban M, Rodriguez D. Endoplasmic reticulum-golgi intermediate compartment membranes and vimentin filaments participate in Vaccinia virus assembly. J Virol. 2002;76(4):1839–55.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Stefanovic S, Windsor M, Nagata KI, Inagaki M, Wileman T. Vimentin rearrangement during African swine fever virus infection involves retrograde transport along microtubules and phosphorylation of vimentin by calcium calmodulin kinase II. J Virol. 2005;79(18):11766–75.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Jaworska J, Coulombe F, Downey J, Tzelepis F, Shalaby K, Tattoli I, et al. NLRX1 prevents mitochondrial induced apoptosis and enhances macrophage antiviral immunity by interacting with influenza virus PB1-F2 protein. Proc Natl Acad Sci U S A. 2014;111(20):E2110–2119.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Jing H, Song T, Cao S, Sun Y, Wang J, Dong W, et al. Nucleotide-binding oligomerization domain-like receptor X1 restricts porcine reproductive and respiratory syndrome virus-2 replication by interacting with viral Nsp9. Virus Res. 2019;268:18–26.

    Article  CAS  PubMed  Google Scholar 

  51. Yang Y, Gibson GE. Succinylation Links metabolism to protein functions. Neurochem Res. 2019;44(10):2346–59.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Huang KY, Hsu JB, Lee TY. Characterization and Identification of Lysine Succinylation Sites based on Deep Learning Method. Sci Rep. 2019;9(1):16175.

    Article  PubMed  PubMed Central  Google Scholar 

  53. El Bakkouri M, Pow A, Mulichak A, Cheung KL, Artz JD, Amani M, et al. The Clp chaperones and proteases of the human malaria parasite Plasmodium Falciparum. J Mol Biol. 2010;404(3):456–77.

    Article  PubMed  Google Scholar 

  54. Malygin AA, Yanshina DD, Karpova GG. Interactions of human ribosomal proteins S16 and S5 with an 18S rRNA fragment containing their binding sites. Biochimie. 2009;91(9):1180–6.

    Article  CAS  PubMed  Google Scholar 

  55. Ian’shina DD, Malygin AA, Karpova GG. [Binding of human ribosomal protein S5 with the 18S rRNA fragment 1203–1236/1521–1698]. Mol Biol (Mosk). 2006;40(3):460–7.

    PubMed  Google Scholar 

  56. Malygin AA, Kossinova OA, Shatsky IN, Karpova GG. HCV IRES interacts with the 18S rRNA to activate the 40S ribosome for subsequent steps of translation initiation. Nucleic Acids Res. 2013;41(18):8706–14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Jheng JR, Wang SC, Jheng CR, Horng JT. Enterovirus 71 induces dsRNA/PKR-dependent cytoplasmic redistribution of GRP78/BiP to promote viral replication. Emerg Microbes Infect. 2016;5:e23.

    CAS  PubMed  PubMed Central  Google Scholar 

  58. Montalbano R, Honrath B, Wissniowski TT, Elxnat M, Roth S, Ocker M, et al. Exogenous hepatitis B virus envelope proteins induce endoplasmic reticulum stress: involvement of cannabinoid axis in liver cancer cells. Oncotarget. 2016;7(15):20312–23.

    Article  PubMed  PubMed Central  Google Scholar 

  59. Fukushima A, Alrob OA, Zhang L, Wagg CS, Altamimi T, Rawat S, et al. Acetylation and succinylation contribute to maturational alterations in energy metabolism in the newborn heart. Am J Physiol Heart Circ Physiol. 2016;311(2):H347–363.

    Article  PubMed  Google Scholar 

  60. Meng X, Dou Y, Cai X. Ultrastructural features of PPRV infection in Vero cells. Virol Sin. 2014;29(5):311–3.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thanked Jingjie PTM BioLab (Hangzhou) Co. Inc for providing technical support.

Funding

This work is financially supported by the Science and Technology Special Projects of Gansu Province (22ZD6NA012-1) and Agricultural Science and Technology Innovation Program (CAAS-ASTIP-2016- LVRI-05).

Author information

Authors and Affiliations

  1. State Key Laboratory for Animal Disease Control and Prevention, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Yanchangpu, Chengguan District, Lanzhou, 730046, Gansu, China

    Xuelian Meng, Xueliang Zhu, Xiangwei Wang & Yuefeng Sun

  2. College of Animal and Veterinary Sciences, Southwest Minzu University, #16, South Section, 1st Ring Road, Chengdu, 610041, Sichuan, China

    Rui Zhang & Zhidong Zhang

Authors
  1. Xuelian Meng
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Xueliang Zhu
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Xiangwei Wang
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Rui Zhang
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Zhidong Zhang
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Yuefeng Sun
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

XLM analyzed and interpreted the proteomic data, and was a major contributor in writing the manuscript. XLZ prepared PPRVvac. XWW helped in preparing figures. RZ prepared samples for LC-MS/MS. ZDZ and YFS helped in writing the manuscript. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Xuelian Meng or Zhidong Zhang.

Ethics declarations

Ethical approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Meng, X., Zhu, X., Wang, X. et al. Comprehensive analysis of the succinylome in Vero cells infected with peste des petits ruminants virus Nigeria 75/1 vaccine strain. BMC Vet Res 21, 45 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12917-025-04496-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12917-025-04496-3

Keywords

BMC Veterinary Research

ISSN: 1746-6148

Contact us