A scalable, GFP-based pipeline for membrane protein overexpression screening and purification

被引：104

作者：

Drew, D

Slotboom, DJ

Friso, G

Reda, T

Genevaux, P

Rapp, M

Meindl-Beinker, NM

Lambert, W

Lerch, M

Daley, DO

Van Wijk, KJ

Hirst, J

Kunji, E

De Gier, JW ^{[1
]}

机构：

[1] Stockholm Univ, Dept Biochem & Biophys, SE-10691 Stockholm, Sweden

[2] MRC, Dunn Human Nutr Unit, Cambridge CB2 2XY, England

[3] Cornell Univ, Dept Plant Biol, Ithaca, NY 14853 USA

[4] Ctr Med, Dept Microbiol & Mol Med, CH-1211 Geneva, Switzerland

来源：

PROTEIN SCIENCE | 2005年 / 14卷 / 08期

基金：

英国医学研究理事会;

关键词：

Escherichia coli; membrane protein overexpression; membrane protein isolation; membrane protein characterization; GFP;

D O I：

10.1110/ps.051466205

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：

We describe a generic, GFP-based pipeline for membrane protein overexpression and purification in Escherichia coli. We exemplify the use of the pipeline by the identification and characterization of E. coli YedZ, a new, membrane-integral flavocytochrome. The approach is scalable and suitable for high-throughput applications. The GFP-based pipeline will facilitate the characterization of the E. coli membrane proteome and serves as an important reference for the characterization of other membrane proteomes.

引用

页码：2011 / 2017

页数：7

共 17 条

[1] Synthesis and bacterial expression of a gene encoding the heme domain of assimilatory nitrate reductase
Barber, MJ
Desai, SK
Marohnic, CC
Hernandez, HH
Pollock, VV
[J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2002, 402 (01) : 38 - 50
[2] BIOCHEMICAL CHANGES IN LIVER ASSOCIATED WITH KWASHIORKOR
BURCH, HB
ARROYAVE, G
SCHWARTZ, R
PADILLA, AM
BEHAR, M
VITERI, F
SCRIMSHAW, NS
[J]. JOURNAL OF CLINICAL INVESTIGATION, 1957, 36 (11) : 1579 - 1587
[3] FLUORIMETRIC ASSAY OF FAD, FMN, AND RIBOFLAVIN
BURCH, HB
[J]. METHODS IN ENZYMOLOGY, 1957, 3 : 960 - 962
[4] Global topology analysis of the Escherichia coli inner membrane proteome
Daley, DO
Rapp, M
Granseth, E
Melén, K
Drew, D
von Heijne, G
[J]. SCIENCE, 2005, 308 (5726) : 1321 - 1323
[5] Rapid topology mapping of Escherichia coli inner-membrane proteins by prediction and PhoA/GFP fusion analysis
Drew, D
Sjöstrand, D
Nilsson, J
Urbig, T
Chin, CN
de Gier, JW
von Heijne, G
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (05) : 2690 - 2695
[6] Assembly and overexpression of membrane proteins in Escherichia coli
Drew, D
Fröderberg, L
Baars, L
de Gier, JWL
[J]. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2003, 1610 (01): : 3 - 10
[7] Green fluorescent protein as an indicator to monitor membrane protein overexpression in Escherichia coli
Drew, DE
von Heijne, G
Nordlund, P
de Gier, JWL
[J]. FEBS LETTERS, 2001, 507 (02) : 220 - 224
[8] Purification and reconstitution of the glutamate carrier GltT of the thermophilic bacterium Bacillus stearothermophilus
Gaillard, I
Slotboom, DJ
Knol, J
Lolkema, JS
Konings, WN
[J]. BIOCHEMISTRY, 1996, 35 (19) : 6150 - 6156
[9] Gobom J, 1999, J MASS SPECTROM, V34, P105, DOI 10.1002/(SICI)1096-9888(199902)34:2<105::AID-JMS768>3.0.CO
[10] 2-4

← 1 2 →