Molecular Diversification Of Peptide Toxins From The Three Kinds Of Wasp

Objective:In this study,the molecular weight of peptides in venoms of Vespa basalis(Smith),Vespa magnifica(Smith)and Vespa mandarinia(Smith)were analyzed by traditional methods.Modern quantitative proteomics was used to quantify the three venom proteins,and the proteins were identified and screened for bioinformatics processing.From the perspective of transcription,the mRNA extracted from the venom glands of V.mandarinia individuals were sequenced by using the second-generation sequencing technology.Methods:1.SDS-PAGE and UPLC-ESI-Q-TOF-MS were used to analyze the relative molecular mass composition of proteins and peptides in the venom of V.basalis,V.magnifica and V.mandarinia.2.Based on the label free protein quantification technique,the relative quantitative analysis of protein enzymatic hydrolysis peptides of venom of V.basalis,V.magnifica and V.mandarinia was carried out by high performance liquid chromatography-mass spectroscopy(LC-MS).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis of unigenes were used to reveale their important functional role in biological processes(BP),molecular functions(MF),cellular components(CC)and metabolic pathway.In addition,the differentially expressed proteins between them were screened and the bioinformatics analysis of these proteins in V.mandarinia vs V.magnifica was performed.3.The mRNA extracted from the venom glands of V.mandarinia wasp individuals was sequenced by using the Illumina HiSeq 2000 high-throughput sequencing technology.Results:1.There were abundant peptides in the venom samples of V.basalis,V.magnifica and V.mandarinia,and the polypeptides and proteins were concentrated respectively in the range of300-3000 Da and 11-48 kDa.2.Through mass spectrometry,39 proteins and 167 peptides were identified by proteomics from V.basalis,V.magnifica and V.mandarinia venom.In which 25 proteins with significant differential in expression were screened by V.mandarinia vs V.magnifica.Of the 39 proteins,nine were up-regulated and sixteen were down-regulated.3.The cDNA libraries from the venom glands of V.mandarinia wasp were sequenced by using the Illumina platform.80,765 clean and high quality unigene sequences were obtained by filtering and assembling.Compared with different databases,it was found that the proteins encoded by transcripts were similar to those of normal cells,and were mainly involved in the process of protein synthesis,secretion,transport and modification.Conclusions:1.This experiment used traditional separation techniques and modern high-throughput sequencing techniques to conduct a relatively systematic study on the molecular weight and composition of venom from V.basalis,V.magnifica and V.mandarinia.From the results,it found that venom was rich in peptides substances,which provided a direction for the selection of follow-up research methods of the venom.2.By combining the mRNA sequencing of V.mandarinia venom glands with the protein mass spectrometry of venom,it was the first time to outline the selection process of peptides and proteins by the venom of V.mandarinia,a signature and highly venomous wasp.