定量蛋白质组学分析链霉素耐药和敏感结核分枝杆菌临床分离株

Quantitative proteomic analysis of streptomycin resistant and sensitive clinical isolates of Mycobacterium tuberculosis

摘要:

摘要:【目的】发现结核分枝杆菌(Mycobacterium tuberculosis)链霉素耐药相关的潜在菌体蛋白。【方法】以结核分枝杆菌临床分离链霉素敏感株01105和结核分枝杆菌H37Rv为对照，采用iTRAQ技术和生物信息学鉴定并相对定量结核分枝杆菌临床分离链霉素耐药株01108菌体蛋白，并通过WEGO功能注释聚类分析01108菌株差异表达蛋白的细胞组分、分子功能和生物进程。【结果】01108菌株分别与01105菌株和H37Rv菌株比较差异表达蛋白为194个和146个，01108菌株与01105菌株和H37Rv比较均差异表达蛋白 121个(共同差异表达蛋白)。差异表达蛋白理论相对分子量和等电点分布广泛，其生物进程主要参与中间代谢、呼吸作用和脂质代谢，分子功能主要为催化活性功能和结合功能。共同差异表达蛋白:7个核糖体蛋白(Rv2785c，Rv0056，Rv0641，Rv0652，Rv0701，Rv1630和Rv2442c)在01108菌株中表达下调;7个蛋白在01108菌株中显著差异表达(上调大于1.20倍或下调小于0.55倍)，分别为巯基过氧化物酶(Rv1932)、酰基载体蛋白脱氢酶(Rv0824c)、30S核糖体蛋白S15(Rv2785c)、丙酮酸脱氢酶E2部分(Rv2215)、双组份转录调控蛋白(Rv3133c)以及假定未知蛋白(Rv2466c和Rv2626c)。【结论】iTRAQ发现了链霉素耐药结核分枝杆菌相对于链霉素敏感结核分枝杆菌和H37Rv共同差异表达蛋白，为进一步探讨结核分枝杆菌链霉素耐药机制奠定了基础。

Abstract:

Abstract:［Objective］To identify specific antigens related to streptomycin resistant (SMr) Mycobacterium tuberculosis．［Methods］Cellular proteins were extracted from SMr clinical isolate 01108，SM-sensitive clinical isolate 01105 and H37Rv． Differential expression proteins were identified with isobaric tags for relative and absolute quantitation (iTRAQ) combined with Nano LC-MS/MS technology．［Results］Approximately 194 and 146 differential expression proteins were identified in 01108 strain compared with the proteomic profiles of 01105 strain and H37Rv，respectively，and 121 proteins were identified in 01108 strain compared with the proteomic profiles of both 01105 strain and H37Rv．Identified proteins showed a pI (isoelectric point) variation between 3.74－12.48 and a molecular mass (Mr) range between 7.63 and 326.2 kDa．Differential expression proteins were mainly associated with metabolism (involved in intermediary metabolism，respiration，and lipid metabolism) and took part in catalysis and binding function．Seven ribosomal proteins (Rv0056，Rv0641，Rv0652，Rv0701，Rv1630，Rv2442c and Rv2785c) and seven proteins (the ratios＞1.20or ＜0.55) were commonly down-regulated in 01108 strain compared with both 01105 strain and H37Rv，i．e．the thiol peroxidase (Rv1932)，acyl carrier protein dehydrogenase (Rv0824c)，30S ribosomal protein S15 (Rv2785c)，acetone acid dehydrogenase E2 part (Rv2215)，two-component transcriptional regulatory protein (Rv3133c) and Hypothetical protein (Rv2466c and Rv2626c)．［Conclusion］Differential expression proteins were found in SMr strain compared with both SM-sensitive strain and H37Rv． Further studies are needed to assess the role of these differential expression proteins in SM resistance．