Protein N-glycosylation in the early stage of Spiroplasma eriocheiris infect Eriocheir sinensis

Abstract

As a novel lethal pathogen of Eriocheir sinensis tremor disease, Spiroplasma eriocheiris, has led into catastrophic economic losses in aquaculture. The hemocytes of E. sinensis is the first target cells of S. eriocheiris. Our study is the first time designed to understanding of the N-glycoproteome dynamics of E. sinensis hemocytes under S. eriocheiris infected at the early phage. In the current study, the N-glycoproteome changes of E. sinensis hemocytes after S. eriocheiris infection were obtained using tandem mass tags (TMT) labeling and affinity enrichment followed by high-resolution Liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) analysis. Using a 1.2-fold change in expression as a physiologically significant benchmark, 167 differentially expressed N-glycosylated proteins were reliably quantified, including 79 up-regulated glycosylated proteins and 88 down-regulated glycosylated proteins subsequented to S. eriocheiris infection. Gene ontology (GO) annotation, protein domain annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation, subcellular localization annotation and protein-protein interaction network were used to analyze those significantly differently expression proteins shown that many biological process and pathway are participate in S. eriocheiris infected host cell, such as phagocytosis, ECM-receptor interaction, lysosome, prophenoloxidase system, and so on. Six selected different expressed glycosylated proteins were studied the transcription at RNA level by qRT-PCR. Our study could serve as a basis to understand the relationship between E. sinensis and the pathogen S. eriocheiris, and also provide reference to study protein N-glycosylation in other crustaceans.