β-桶外膜蛋白质生成机制的活细胞探究  

Exploring the Biogenesis Mechanism ofβ-Barrel Outer Membrane Proteins in Living Cells

作　　者：王妍 昌增益[2] WANG Yan;CHANG Zeng-Yi(Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation,College of Life Sciences,Fujian Normal University,Fuzhou 350108,China;State Key Laboratory of Protein and Plant Gene Research,School of Life Sciences,Center for Protein Science,Peking University,Beijing 100871,China)

机构地区：[1]细胞逆境响应与代谢调控福建省高校重点实验室,福建师范大学生命科学学院,福州350108 [2]蛋白质与植物基因研究国家重点实验室,北京大学生命科学学院,北京大学蛋白质科学中心,北京100871

出　　处：《中国生物化学与分子生物学报》2025年第1期36-51,共16页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金委原始创新探索项目(No.32150003);国家自然科学基金面上项目(No.3215000010,31971189,49931670775,31470766);启东-北大生命科学学院创新基金资助。

摘　　要：β-桶外膜蛋白是革兰氏阴性菌外膜以及真核细胞线粒体和叶绿体外膜上广泛存在的一类膜整合蛋白质。细菌β-桶外膜蛋白质的体内生成是一个十分复杂的过程,包括多肽链在细胞质内的合成,跨内膜转运,穿过亲水的膜间质空间最后到达外膜,完成最后的折叠。在β-桶外膜蛋白生成过程中,膜间质中的质量控制因子SurA、Skp和DegP等通过何种机制帮助β-桶外膜蛋白在亲水的膜间质环境中完成运输和折叠,以及内膜上的SecYEG-SecDF-YidC复合体和外膜上的BAM复合体以何种形式与膜间质质量控制因子相互作用,并共同完成β-桶外膜蛋白的体内合成,是一直以来亟待回答的科学问题。本文就革兰氏阴性菌β-桶外膜蛋白生成机制的研究进展,以及各种新技术尤其是非天然氨基酸体内光交联技术,及高通量聚丙烯酰胺凝胶电泳(SDS-PAGE)技术在研究β-桶外膜蛋白折叠中间体的应用进行介绍,为外膜蛋白生成机制的研究提供新的思路。The β-barrel outer membrane protein is a kind of membrane integration protein widely existing in the outer membranes of Gram-negative bacteria,mitochondria and chloroplasts.The biogenesis of β-barrel outer membrane proteins in living bacterial cells is an extremely complex process,including the synthesis of nascent polypeptide chains in the cytoplasm,the translocation across the inner membrane,the transportation through the hydrophilic periplasmic space and finally the folding,assembly and insertion to the outer membrane to complete the whole process.During the biogenesis of β-barrel outer membrane proteins,it is critical to unveil how the periplasmic quality control factors such as SurA,Skp and DegP assist the folding and transportation of the nascent β-barrel outer membrane proteins through the hydrophilic periplasmic space,and how the SecYEG-SecDF-Yidc complex on the inner membrane and the BAM complex on the outer membrane act during the biogenesis of β-barrel outer membrane proteins.This review delves into the research progress of the mechanism of β-barrel outer membrane protein biogenesis in Gram-negative bacteria and also the application of various new technologies such as in vivo protein photo-crosslinking analysis and high through-put protein electrophoresis(SDS-PAGE)for exploring such dynamic cellular processes as protein biogenesis.

关 键 词：β-桶外膜蛋白 膜蛋白生成 非天然氨基酸 体内蛋白质光交联 蛋白质超级复合体 

分 类 号：Q71[生物学—分子生物学]