A general strategy for the expression of bacterial membrane transport and receptor genes in Escherichia coli is described. Expression is amplified so that the encoded proteins comprise 5–35% of E. coli inner membrane protein. Depending upon their topology, proteins are produced with RGSH6 or a Strep tag at the C-terminus. These enable purification in mg quantities for crystallization and NMR studies. Examples of one nutrient uptake and one multidrug extrusion protein from Helicobacter pylori are described. This strategy is successful for membrane proteins from H. pylori, E. coli, Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Microbacterium liquefaciens, Brucella abortus, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides, Streptomyces coelicolor and Rhodobacter sphaeroides.
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August 2005
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Conference Article|
August 01 2005
Active membrane transport and receptor proteins from bacteria
M. Saidijam;
M. Saidijam
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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K.E. Bettaney;
K.E. Bettaney
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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G. Szakonyi;
G. Szakonyi
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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G. Psakis;
G. Psakis
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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K. Shibayama;
K. Shibayama
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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S. Suzuki;
S. Suzuki
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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J.L. Clough;
J.L. Clough
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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V. Blessie;
V. Blessie
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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A. Abu-bakr;
A. Abu-bakr
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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S. Baumberg;
S. Baumberg
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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J. Meuller;
J. Meuller
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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C.K. Hoyle;
C.K. Hoyle
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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S.L. Palmer;
S.L. Palmer
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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P. Butaye;
P. Butaye
†Department of Bacteriology and Immunology, Veterinary and Agrochemical Research Centre, VAR-CODA-CERVA, Groeselenberg 99, B-1180 Ukkel, Belgium
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K. Walravens;
K. Walravens
†Department of Bacteriology and Immunology, Veterinary and Agrochemical Research Centre, VAR-CODA-CERVA, Groeselenberg 99, B-1180 Ukkel, Belgium
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S.G. Patching;
S.G. Patching
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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J. O'Reilly;
J. O'Reilly
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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N.G. Rutherford;
N.G. Rutherford
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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R.M. Bill;
R.M. Bill
‡Aston Academy of Life Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, U.K.
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D.I. Roper;
D.I. Roper
§Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV47AL, U.K.
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M.K. Phillips-Jones;
M.K. Phillips-Jones
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
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P.J.F. Henderson
P.J.F. Henderson
1
*Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
1To whom correspondence should be addressed (email p.j.f.henderson@leeds.ac.uk).
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Publisher: Portland Press Ltd
Received:
April 04 2005
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2005 The Biochemical Society
2005
Biochem Soc Trans (2005) 33 (4): 867–872.
Article history
Received:
April 04 2005
Citation
M. Saidijam, K.E. Bettaney, G. Szakonyi, G. Psakis, K. Shibayama, S. Suzuki, J.L. Clough, V. Blessie, A. Abu-bakr, S. Baumberg, J. Meuller, C.K. Hoyle, S.L. Palmer, P. Butaye, K. Walravens, S.G. Patching, J. O'Reilly, N.G. Rutherford, R.M. Bill, D.I. Roper, M.K. Phillips-Jones, P.J.F. Henderson; Active membrane transport and receptor proteins from bacteria. Biochem Soc Trans 1 August 2005; 33 (4): 867–872. doi: https://doi.org/10.1042/BST0330867
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