Differentially Proteins in Colaphellus bowringi Treated with Sip Toxin Protein by iTRAQ Quantitative Proteomics
YIN Yan-Li1, SHA Jun-Xue1, ZHANG You-Ran1, TAN Jia-Li1, SUN Jing1, GAO Ze-Ping1, LI Hai-Tao1,2,*
1 College of Life Science, Northeast Agricultural University, Harbin 150030, China; 2 Institute of Plant Protection, Chinese Academy of Agricultural Sciences / State Key Laboratory of Biology of Plant Diseases and Pests, Beijing 100193, China
Abstract：Secreted insecticidal protein (Sip) is a secretory insecticidal protein of Bacillus thuringiensis (Bt) insecticidal family, which is toxic to Coleoptera larvae. Colaphellus bowringi is a common pest of Cruciferae. Sip protein has high insecticidal activity against the Colaphellus bowringi. In this experiment, the isotope relative labeling and absolute quantification technique (iTRAQ) was used to analyze the differential expression protein of Colaphellus bowringi protein, and the differential proteins were screened and subjected to functional annotation and enrichment analysis. Quantitative analysis was performed by ProteinpilotTM V4.5 software, and Gene Ontology (GO) function annotation, Cluster of Orthologous Groups of proteins (COG)function annotation and KEGG analysis were performed, and functional enrichment analysis (GO, KEGG enrichment) was performed on all significantly differential proteins. Compared with the control group, 47 proteins were identified in the experimental group with quantitative information, including 27 up-regulated proteins and 20 down-regulated proteins. Through GO function annotation, COG and KEGG analysis, it was found that these differential proteins mainly involved molecular functions such as binding and catalytic activity, among which the functions unique to up-regulated proteins were electron carrier activity, antioxidant activity and molecular converter activity. These proteins mainly performed biological processes such as cell processes and metabolic processes, in which only up-regulated proteins were involved in death, immune system processes, exercise, and viral replication. These proteins mainly consisted of cells, cell parts, organelles, etc. It was predicted that these differential proteins might have functions such as post-translational modification, protein turnover, chaperone, translation, ribosome structure and biosynthesis, translation, ribosome structure and biosynthesis, etc. In the biochemical metabolic pathway and signal transduction pathway, the up-regulated protein and the down-regulated protein shared a common pathway for the biosynthesis of secondary metabolites. The pathways that were important for up-regulating differential proteins were metabolic pathways, microbial metabolism in different environments, secondary metabolic biosynthesis, and pyruvate metabolism. The specific pathways for down-regulating proteins included the ribosomal pathway, systemic lupus erythematosus, and pancreatic secretory pathway. Through enrichment analysis, the main biological functions of these differential proteins were found to be nuclear nucleosomes, non-membrane bounded organelles, intracellular non-membrane bounded organelles, nucleosomes, ribosomes, COPI envelope vesicles, protein-DNA complex, secretory granules, nuclear chromatin, Golgi heap, etc. After treatment with Sip toxin protein, some receptors common to Bt toxin were found to be differentially expressed, among which alkaline phosphatase (Alp), G-protein, cadherin and aminopeptidase N (APN) were differentially expressed. The results provide a reference for the study of the mechanism and for the proteomics study of Sip toxin.
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