In Situ Structural Characterization of a Recombinant Protein in Native Escherichia coli Membranes with Solid-State Magic-Angle-Spinning NMR

被引：71

作者：

Fu, Riqiang ^{[2
]}

Wang, Xingsheng ^{[1
]}

Li, Conggang ^{[3
]}

Santiago-Miranda, Adriana N. ^{[4
]}

Pielak, Gary J. ^{[5
]}

Tian, Fang ^{[1
]}

机构：

[1] Penn State Univ, Dept Biochem & Mol Biol, Hershey, PA 17033 USA

[2] Natl High Magnet Field Lab, Tallahassee, FL 32310 USA

[3] Chinese Acad Sci, Wuhan Inst Phys & Math, State Key Lab Magnet Resonance & Atom & Mol Phys, Wuhan 430071, Peoples R China

[4] Univ Puerto Rico, Dept Chem Engn, Mayaguez, PR 00681 USA

[5] Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2011年 / 133卷 / 32期

基金：

中国国家自然科学基金; 美国国家科学基金会; 美国国家卫生研究院;

关键词：

AMYLOID PRECURSOR PROTEIN; RESONANCE ASSIGNMENT; ALZHEIMER-DISEASE; SECONDARY STRUCTURE; PURPLE MEMBRANES; LIPID-MEMBRANES; COAT PROTEIN; C-13; BACTERIORHODOPSIN; SPECTROSCOPY;

D O I：

10.1021/ja204062v

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The feasibility of using solid-state magic-angle-spinning NMR spectroscopy for in situ structural characterization of the LR11 (sorLA) transmembrane domain (TM) in native Escherichia coli membranes is presented. LR11 interacts with the human amyloid precursor protein (APP), a central player in the pathology of Alzheimer's disease. The background signals from E. coli lipids and membrane proteins had only minor effects on the LR11 TM resonances. Approximately 50% of the LR11 TM residues were assigned by using C-13 PARIS data. These assignments allowed comparisons of the secondary structure of the LR11 TM in native membrane environments and commonly used membrane mimics (e.g., micelles). In situ spectroscopy bypasses several obstacles in the preparation of membrane proteins for structural analysis and offers the opportunity to investigate how membrane heterogeneity, bilayer asymmetry, chemical gradients, and macromolecular crowding affect the protein structure.

引用

页码：12370 / 12373

页数：4