Protein solubility and folding monitored in vivo by structural complementation of a genetic marker protein

被引：132
作者：
Wigley, WC
Stidham, RD
Smith, NM
Hunt, JF
Thomas, PJ
机构：
[1] Univ Texas, SW Med Ctr, Dept Physiol, Dallas, TX 75235 USA
[2] Univ Texas, SW Med Ctr, Grad Program Mol Biophys, Dallas, TX 75235 USA
[3] Columbia Univ, Dept Biol Sci, New York, NY 10027 USA
来源：
NATURE BIOTECHNOLOGY | 2001年 / 19卷 / 02期
关键词：
aggregation; folding; solubility; complementation; protein-folding diseases;
D O I：
10.1038/84389
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Protein misfolding is the basis of a number of human diseases and presents an obstacle to the production of soluble recombinant proteins. We present a general method to assess the solubility and folding of proteins in vivo. The basis of this assay is structural complementation between the alpha- and omega -fragments of beta -galactosidase (beta -gal). Fusions of the alpha -fragment to the C terminus of target proteins with widely varying in vivo folding yield and/or solubility levels, including the Alzheimer's amyloid beta (A beta) peptide and a non-amyloidogenic mutant thereof, reveal an unambiguous correlation between beta -gal activity and the solubility/folding of the target. Thus, structural complementation provides a means of monitoring protein solubility/misfolding in vivo, and should find utility in the screening for compounds that influence the pathological consequences of these processes.
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收藏
页码：131 / 136
页数：6
相关论文
共 59 条
[11]   Androgen receptor mutation in Kennedy's disease [J].
Fischbeck, KH ;
Lieberman, A ;
Bailey, CK ;
Abel, A ;
Merry, DE .
PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES, 1999, 354 (1386) :1075-1078
[12]   Pharmacological rescue of mutant p53 conformation and function [J].
Foster, BA ;
Coffey, HA ;
Morin, MJ ;
Rastinejad, F .
SCIENCE, 1999, 286 (5449) :2507-2510
[13]   Axon pathology in Parkinson's disease and Lewy body dementia hippocampus contains α-, β-, and γ-synuclein [J].
Galvin, JE ;
Uryu, K ;
Lee, VMY ;
Trojanowski, JQ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (23) :13450-13455
[14]   INTERACTION OF HEAT-SHOCK PROTEIN-70 WITH P53 TRANSLATED INVITRO - EVIDENCE FOR INTERACTION WITH DIMERIC P53 AND FOR A ROLE IN THE REGULATION OF P53 CONFORMATION [J].
HAINAUT, P ;
MILNER, J .
EMBO JOURNAL, 1992, 11 (10) :3513-3520
[15]   Models of amyloid seeding in Alzheimier's disease and scrapie: Mechanistic truths and physiological consequences of the time-dependent solubility of amyloid proteins [J].
Harper, JD ;
Lansbury, PT .
ANNUAL REVIEW OF BIOCHEMISTRY, 1997, 66 :385-407
[16]   DEMONSTRATION OF AMYLOID-A (AA) PROTEIN AND AMYLOID-P COMPONENT (AP) IN DEPOSITS OF SYSTEMIC AMYLOIDOSIS ASSOCIATED WITH RENAL ADENOCARCINOMA [J].
HIND, CRK ;
TENNENT, GA ;
EVANS, DJ ;
PEPYS, MB .
JOURNAL OF PATHOLOGY, 1983, 139 (02) :159-166
[17]   Identification of in vivo substrates of the chaperonin GroEL [J].
Houry, WA ;
Frishman, D ;
Eckerskorn, C ;
Lottspeich, F ;
Hartl, FU .
NATURE, 1999, 402 (6758) :147-154
[18]   NMR structure and mutagenesis of the Fas (APO-1/CD95) death domain [J].
Huang, BH ;
Eberstadt, M ;
Olejniczak, ET ;
Meadows, RP ;
Fesik, SW .
NATURE, 1996, 384 (6610) :638-641
[19]   Overproduction, in Escherichia coli, of soluble taxadiene synthase, a key enzyme in the taxol biosynthetic pathway [J].
Huang, EX ;
Huang, QL ;
Wildung, MR ;
Croteau, R ;
Scott, AI .
PROTEIN EXPRESSION AND PURIFICATION, 1998, 13 (01) :90-96
[20]   Crystal structure of the ATP-binding subunit of an ABC transporter [J].
Hung, LW ;
Wang, IXY ;
Nikaido, K ;
Liu, PQ ;
Ames, GFL ;
Kim, SH .
NATURE, 1998, 396 (6712) :703-707
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