Optimization of membrane protein overexpression and purification using GFP fusions

Nature Methods

Volumn 3, Issue 4, 2006, Pages 303-313

  Drew, David ^a   Lerch, Mirjam ^a   Kunji, Edmund ^b   Slotboom, Dirk Jan ^c   de Gier, Jan Willem ^a  

^a STOCKHOLM UNIVERSITY   (Sweden)

^b UNIVERSITY OF CAMBRIDGE   (United Kingdom)

^c UNIVERSITY OF GRONINGEN   (Netherlands)

Author keywords

[No Author keywords available]

Indexed keywords

GREEN FLUORESCENT PROTEIN; HYBRID PROTEIN; MEMBRANE PROTEIN;

ARTICLE; BACTERIAL STRAIN; CARBOXY TERMINAL SEQUENCE; CELL CULTURE; CELL SUSPENSION; CELLULAR DISTRIBUTION; CENTRIFUGATION; ESCHERICHIA COLI; FLUORESCENCE ANALYSIS; IMMUNOBLOTTING; NONHUMAN; PREDICTION; PRIORITY JOURNAL; PROCESS OPTIMIZATION; PROTEIN ANALYSIS; PROTEIN EXPRESSION; PROTEIN FUNCTION; PROTEIN LOCALIZATION; PROTEIN PURIFICATION; PROTEIN STRUCTURE;

ESCHERICHIA COLI PROTEINS; GENE EXPRESSION REGULATION, BACTERIAL; GREEN FLUORESCENT PROTEINS; LUMINESCENT AGENTS; MEMBRANE PROTEINS; RECOMBINANT FUSION PROTEINS;

BACTERIA (MICROORGANISMS); ESCHERICHIA COLI;

EID: 33645454462     PISSN: 15487091     EISSN: 15491676     Source Type: Journal    
DOI: 10.1038/nmeth0406-303     Document Type: Article

References (13)

1. Drew, D.E., von Heijne, G., Nordlund, P. & de Gier, J.W. Green fluorescent protein as an indicator to monitor membrane protein overexpression in Escherichia coli. FEBS Lett. 507, 220-224 (2001).
2. Drew, D. et al. Rapid topology mapping of Escherichia coli inner-membrane proteins by prediction and PhoA/GFP fusion analysis. Proc. Natl. Acad. Sci. USA 99, 2690-2695 (2002).
3. Drew, D. et al. A scalable, GFP-based pipeline for membrane protein overexpression screening and purification. Protein Sci. 14, 2011-2017 (2005).
4. Pedelacq, J.D., Cabantous, S., Tran, T., Terwilliger, T.C. & Waldo, G.S. Engineering and characterization of a superfolder green fluorescent protein. Nat. Biotechnol. 24, 79-88 (2006).
5. Wallin, E. & von Heijne, G. Genome-wide analysis of integral membrane proteins from eubacterial, archaean, and eukaryotic organisms. Protein Sci. 7, 1029-1038 (1998).
6. Granseth, E., Daley, D.O., Rapp, M., Melen, K. & von Heijne, G. Experimentally constrained topology models for 51,208 bacterial inner membrane proteins. J. Mol. Biol. 352, 489-494 (2005).
7. Daley, D.O. et al. Global topology analysis of the Escherichia coli inner membrane proteome. Science 308, 1321-1323 (2005).
8. Rapp, M. et al. Experimentally based topology models for E. coli inner membrane proteins. Protein Sci. 13, 937-945 (2004).
9. Wang, D.N. et al. Practical aspects of overexpressing bacterial secondary membrane transportes for structural studies. Biochim. Biophys. Acta 1610, 23-36 (2003).
10. Vethanayagam, J.G. & Flower, A.M. Decreased gene expression from T7 promoters may be due to impaired production of active T7 RNA polymerase. Microb. Cell Fact. 4, 3 (2005).
11. Feilmeier, B.J., Iseminger, G., Schroeder, D., Webber, H. & Phillips, G.J. Green fluorescent protein functions as a reporter for protein localization in Escherichia coli. J. Bacteriol. 182, 4068-4076 (2000).
12. Sarramegna, V., Talmont, F., Seree de Roch, M., Milon, A. & Demange, P. Green fluorescent protein as a reporter of human mu-opioid receptor overexpression and localization in the methylotrophic yeast Pichia pastoris. J. Biotechnol. 99, 23-39 (2002).
13. Huang, Y., Lemieux, M.J., Song, J., Auer, M. & Wang, D.N. Structure and mechanism of the glycerol-3-phosphate transporter from Escherichia coli. Science 301, 616-620 (2003).