Effects of Recombinant Protein Expression on Green Fluorescent Protein Diffusion in Escherichia coli

被引：27

作者：

Slade, Kristin M. ^{[1
]}

Baker, Rachael ^{[2
]}

Chua, Michael ^{[3
]}

Thompson, Nancy L. ^{[1
]}

Pielak, Gary J. ^{[1
,2
,4
]}

机构：

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

[2] Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USA

[3] Univ N Carolina, Dept Cell & Mol Physiol, Chapel Hill, NC 27599 USA

[4] Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USA

来源：

BIOCHEMISTRY | 2009年 / 48卷 / 23期

关键词：

CELL NMR-SPECTROSCOPY; ALPHA-SYNUCLEIN; IN-VIVO; PHOTOBLEACHING RECOVERY; OSMOTIC-STRESS; LIVING CELLS; VOLUME; CONSEQUENCES; MOBILITY; CONFINEMENT;

D O I：

10.1021/bi9004107

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Fluorescence recovery after photobleaching was used to measure the diffusion coefficient of green fluorescent protein (GFP, 27 kDa) in Escherichia coli in the presence or absence of four coexpressed proteins: cytoplasmic maltose binding protein (42 kDa), tau-40 (45 kDa), alpha-synuclein (14 kDa), or calmodulin (17 kDa). The G FP diffusion coefficient remains constant regardless of the type of coexpresseed protein and whether or not the coexpressed protein was induced. We conclude that expression of these soluble proteins has little to no effect oil the diffusion of GFP. These results have implications for the utility of in-cell nuclear magnetic resonance spectroscopy.

引用

页码：5083 / 5089

页数：7

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