耐碳青霉烯类肺炎克雷伯菌耐药机制的定量蛋白质组分析

Analysis of Quantitative Proteome in Drug Resistance Mechanism of Carbapenem-resistant Klebsiella pneumoniae

为了寻找耐碳青霉烯类肺炎克雷伯菌耐药相关蛋白,本研究通过提取2017年莆田学院附属医院分离的3株耐碳青霉烯类肺炎克雷伯菌耐药株和2株耐碳青霉烯类肺炎克雷伯菌敏感株的总蛋白,采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)技术合并液相色谱-质谱联用技术(liquid chromatography-mass spectrometry,LC-MS/MS)分析和鉴定蛋白表达图谱,筛选差异表达蛋白,并对显著性差异蛋白进行GO功能及KEGG通路富集分析。我们共鉴定出3117个蛋白,定量检测到的蛋白有2920个,其中538个蛋白有显著性差异。GO功能富集分析发现,与肺炎克雷伯菌耐药相关的蛋白有β-内酰胺酶、青霉素结合蛋白、修饰氨基糖苷类的钝化酶、金属肽酶及其它相关蛋白。对β-内酰胺相关蛋白KEGG通路富集分析,涉及β-内酰胺酶活性的KPC型和SHV型基因得到验证。本研究为耐碳青霉烯类肺炎克雷伯菌耐药机制提供了理论依据。

This study aims to search for drug resistance-related proteins of Klebsiella pneumoniae.Three carbapenem-resistant K.pneumoniae and two carbapenem-sensitive strains were isolated from the Affiliated Hospital of Putian University in 2017 and made into protein samples.iTRAQ labeling and LC-MS/MS were performed to identify the total number and quantitative determination of proteins,respectively.Gene ontology functional and KEGG pathway enrichment analyses were performed for all the proteins with significant differences.A total of3117 proteins were identified in this project.Among them,2920 proteins had quantitative,and 538 proteins with significant differences were screened.Significant differences were observed in GO functional classification and concentration.Proteins,including beta-lactamase,penicillin-binding protein,aminoglycoside-modified inactivating enzyme,metalloproteinase,and related proteins,were found to be related to the drug resistance of K.pneumoniae.KEGG pathway enrichment analysis of beta-lactam-related proteins was conducted.KPC and SHV genes related to beta-lactamase activity were identified.This study provides valuable reference for future studies on drug resistance mechanisms of K.pneumoniae.