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Proteomics and Phosphoproteomic Analysis to Identify Spleen of Takifugu rubripes Infected Cryptocaryon irritans

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Abstract

Takifugu rubripes is important commercially fish species in China and it is under serious threat from white spot disease (cyptocaryoniasis), which leads to heavy economic losses. In this study, we used proteomics and phosphoproteomic analysis to identify differentially abundant proteins in the spleen of T. rubripes infected with the Cryptocaryon irritans. We identified 5,307 proteins and 6,644 phosphorylated sites on 2,815 phosphoproteins using high-throughput proteomics analysis of the spleen of T. rubripes based on 26,421 unique peptides and 5,013 modified peptides, respectively. The 5,307 quantified host proteins, 40 were upregulated and 43 were downregulated in the infection group compared to the control group. Among the 2815 phosphoproteins, 44/120 were upregulated/downregulated, and 62/151 were upregulated/downregulated in the 6644 quantified phosphosites. Using the combination of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses, screening for significantly different phosphoproteins, motif analysis and protein–protein interaction analysis, we ultimately identified three phosphorylated proteins (G-protein-signaling modulator 1-like, zinc finger protein 850-like, and histone H1-like) and three phosphorylated protein kinases (serine/threonine-protein kinase homolog isoform X2, mitogen-activated protein kinase 5-like, and protein kinase C theta type) as potential biomarkers for T. rubripes immune responses. We then screened the phosphorylation sites of these biomarker proteins for further verification. Based on our results, we speculate that phosphorylation modification of the phosphorylation sites is involved in the immunity of T. rubripes against C. irritans.

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Data Availability

I have shared the link to the manuscript and Proteomics data.The datasets generated and/or analysed during the current study are available in the [PRIDE] repository, [http://www.ebi.ac.uk/pride ID: PXD034249 Username: reviewer_pxd034249@ebi.ac.uk, Password: ooxsMcKB].

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Acknowledgements

We thank our colleagues who contributed to this study. We thank the Hangzhou PTM bioinformatics Team (Hangzhou, China) for providing proteomics services. This work was supported by grants from Tackling-key Project of Education Department Funding for scientific research of Liaoning Province (ZL202002), and Technology Major Special Plan Project of Liaoning Province (2020JH1/10200002).

Funding

This work was supported by grants from Tackling-key Project of Education Department Funding for scientific research of Liaoning Province (ZL202002), and Technology Major Special Plan Project of Liaoning Province (2020JH1/10200002).

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Authors and Affiliations

  1. College of Life Science, Liaoning Normal University, Dalian, 116081, China

    Jianxin Cheng

  2. College of Marine Technology and Environment, Dalian Ocean University, No.52, Heishijiao Road, Dalian, 116023, China

    Cheng Zhou, Xiaohao Li & Pengfei Liu

  3. Key Laboratory of Environment Controlled Aquaculture (KLECA), Ministry of Education, 52 Heishijiao Street, Dalian, 116023, China

    Jianxin Cheng, Yuqing Xia, Cheng Zhou, Xiaohao Li & Pengfei Liu

  4. School of Marine Sciences, Ningbo University, Ningbo, Zhejiang, 315211, People’s Republic of China

    Yuqing Xia

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Contributions

Jianxin Cheng: Conception and design, data analysis, Writing-original draft; Conceptualization, writing this manuscript; Yuqing Xia: Data analysis; Cheng Zhou and Xiaohao Li: Acquisition test material; Pengfei Liu: Conceptualization, Supervision, Funding acquisition. All authors have given approval to the final version of the manuscript, decided to submit the work for publication.

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Correspondence to Pengfei Liu.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. All fish in this study were handled in strict accordance with China legislation on scientific procedures on living animals. All protocols involving fish were approved by the Institutional Animal Care and Use Committees at Dalian Ocean University.

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Cheng, J., Xia, Y., Zhou, C. et al. Proteomics and Phosphoproteomic Analysis to Identify Spleen of Takifugu rubripes Infected Cryptocaryon irritans. Mar Biotechnol 25, 291–313 (2023). https://doi.org/10.1007/s10126-023-10205-8

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