Overexpression of mammalian integral membrane proteins for structural studies

被引:139
作者
Tate, CG [1 ]
机构
[1] MRC, Mol Biol Lab, Cambridge CB2 2QH, England
关键词
structure; crystallization; serotonin transporter;
D O I
10.1016/S0014-5793(01)02711-9
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
Recent successes in the determination of atomic resolution structures of integral membrane proteins have relied on purifying the proteins from abundant natural sources. In contrast, the majority of mammalian receptors, ion channels and transporters need to be overexpressed to obtain sufficient material for structural studies. This has often proved to be very difficult. Overexpression studies on a wide range of mammalian membrane proteins have shown that a few can be expressed functionally in bacteria, but many others require an insect or mammalian cell host for activity or high level expression. The serotonin transporter, which has been expressed in all the major hosts available, is a good example that has given insights into the problem of overexpressing mammalian membrane proteins for structural studies. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:94 / 98
页数:5
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  • [1] TRANSLATION RATES AND MISREADING CHARACTERISTICS OF RPSD MUTANTS IN ESCHERICHIA-COLI
    ANDERSSON, DI
    BOHMAN, K
    ISAKSSON, LA
    KURLAND, CG
    [J]. MOLECULAR AND GENERAL GENETICS, 1982, 187 (03): : 467 - 472
  • [2] Three-dimensional map of the plasma membrane H+-ATPase in the open conformation
    Auer, M
    Scarborough, GA
    Kühlbrandt, W
    [J]. NATURE, 1998, 392 (6678) : 840 - 843
  • [3] BAKER C, 1999, THESIS TRINITY COLL
  • [4] A temperature-regulated replicon-based DNA expression system
    Boorsma, M
    Nieba, L
    Koller, D
    Bachmann, MF
    Bailey, JE
    Renner, WA
    [J]. NATURE BIOTECHNOLOGY, 2000, 18 (04) : 429 - 432
  • [5] Structure of the MscL homolog from Mycobacterium tuberculosis:: A gated mechanosensitive ion channel
    Chang, G
    Spencer, RH
    Lee, AT
    Barclay, MT
    Rees, DC
    [J]. SCIENCE, 1998, 282 (5397) : 2220 - 2226
  • [6] External cysteine residues in the serotonin transporter
    Chen, JG
    LiuChen, S
    Rudnick, G
    [J]. BIOCHEMISTRY, 1997, 36 (06) : 1479 - 1486
  • [8] The structure of the potassium channel:: Molecular basis of K+ conduction and selectivity
    Doyle, DA
    Cabral, JM
    Pfuetzner, RA
    Kuo, AL
    Gulbis, JM
    Cohen, SL
    Chait, BT
    MacKinnon, R
    [J]. SCIENCE, 1998, 280 (5360) : 69 - 77
  • [9] Structure of a glycerol-conducting channel and the basis for its selectivity
    Fu, DX
    Libson, A
    Miercke, LJW
    Weitzman, C
    Nollert, P
    Krucinski, J
    Stroud, RM
    [J]. SCIENCE, 2000, 290 (5491) : 481 - 486
  • [10] OVEREXPRESSION OF INTEGRAL MEMBRANE-PROTEINS FOR STRUCTURAL STUDIES
    GRISSHAMMER, R
    TATE, CG
    [J]. QUARTERLY REVIEWS OF BIOPHYSICS, 1995, 28 (03) : 315 - 422