Functional Reconstitution of Staphylococcus aureus Truncated AgrC Histidine Kinase in a Model Membrane System

	Functional Reconstitution of Staphylococcus aureus Truncated AgrC Histidine Kinase in a Model Membrane System
	Wang, Lina 1; Quan, Chunshan 2,3; Liu, Baoquan 2,3; Wang, Jianfeng 2,3; Xiong, Wen 2,3; Zhao, Pengchao 1; Fan, Shengdi 2,3
刊名	plos one
	2013-11-26
卷号	8 期号:11
英文摘要	the integral membrane protein agrc is a histidine kinase whose sensor domains interact with an autoinducing peptide, resulting in a series of downstream responses. in this study, truncated agrc(tm5-6c) and agrc(tm5-6c)-gfp with gfp as a reporter gene were produced using a bacterial system. purified agrc(tm5-6c) and agrc(tm5-6c)-gfp were reconstituted into liposomes by a detergent-mediated method. to achieve high-yield protein incorporation, we investigated the effect of different detergents on protein reconstitution efficiency. the highest incorporation was found with n, n-dimethyldode-cylamine noxide during complete liposome solubilization, which resulted in a yield of 85 +/- 5%. the cooh-terminus of the protein agrc(tm5-6c) was almost exclusively oriented towards the inside of the vesicles. agrc(tm5-6c) in proteoliposomes exhibited approximately a 6-fold increase in constitutive activity compared with agrc(tm5-6c) in detergent micelles. the reconstitution of agrc(tm5-6c) or agrc(tm5-6c)-gfp was characterized using dynamic light scattering, fluorescence microscopy, and transmission electron microscopy. based on the results, the optimal conditions for protein incorporation were defined. these findings contribute to the study of membrane protein structure and function in vitro using a reconstitution system.
WOS标题词	science & technology
类目[WOS]	multidisciplinary sciences
研究领域[WOS]	science & technology - other topics
关键词[WOS]	size-exclusion chromatography ; escherichia-coli ; 2-dimensional crystallization ; protein reconstitution ; autoinducing peptides ; receptor ; sensor ; detergents ; liposomes ; virulence
收录类别	SCI
语种	英语
WOS记录号	WOS:000327546400027
公开日期	2015-11-10
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/137718]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian, Peoples R China 2.Dalian Nationalities Univ, Dept Life Sci, Dalian, Peoples R China 3.State Ethn Affairs Commiss, Minist Educ, Dalian, Peoples R China
推荐引用方式 GB/T 7714	Wang, Lina,Quan, Chunshan,Liu, Baoquan,et al. Functional Reconstitution of Staphylococcus aureus Truncated AgrC Histidine Kinase in a Model Membrane System[J]. plos one,2013,8(11).
APA	Wang, Lina.,Quan, Chunshan.,Liu, Baoquan.,Wang, Jianfeng.,Xiong, Wen.,...&Fan, Shengdi.(2013).Functional Reconstitution of Staphylococcus aureus Truncated AgrC Histidine Kinase in a Model Membrane System.plos one,8(11).
MLA	Wang, Lina,et al."Functional Reconstitution of Staphylococcus aureus Truncated AgrC Histidine Kinase in a Model Membrane System".plos one 8.11(2013).