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.

引用

页码：131 / 136

页数：6

共 59 条

[11] Androgen receptor mutation in Kennedy's disease
Fischbeck, KH
Lieberman, A
Bailey, CK
Abel, A
Merry, DE
[J]. PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES, 1999, 354 (1386) : 1075 - 1078
[12] Pharmacological rescue of mutant p53 conformation and function
Foster, BA
Coffey, HA
Morin, MJ
Rastinejad, F
[J]. SCIENCE, 1999, 286 (5449) : 2507 - 2510
[13] Axon pathology in Parkinson's disease and Lewy body dementia hippocampus contains α-, β-, and γ-synuclein
Galvin, JE
Uryu, K
Lee, VMY
Trojanowski, JQ
[J]. 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
HAINAUT, P
MILNER, J
[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
Harper, JD
Lansbury, PT
[J]. 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
HIND, CRK
TENNENT, GA
EVANS, DJ
PEPYS, MB
[J]. JOURNAL OF PATHOLOGY, 1983, 139 (02) : 159 - 166
[17] Identification of in vivo substrates of the chaperonin GroEL
Houry, WA
Frishman, D
Eckerskorn, C
Lottspeich, F
Hartl, FU
[J]. NATURE, 1999, 402 (6758) : 147 - 154
[18] NMR structure and mutagenesis of the Fas (APO-1/CD95) death domain
Huang, BH
Eberstadt, M
Olejniczak, ET
Meadows, RP
Fesik, SW
[J]. NATURE, 1996, 384 (6610) : 638 - 641
[19] Overproduction, in Escherichia coli, of soluble taxadiene synthase, a key enzyme in the taxol biosynthetic pathway
Huang, EX
Huang, QL
Wildung, MR
Croteau, R
Scott, AI
[J]. PROTEIN EXPRESSION AND PURIFICATION, 1998, 13 (01) : 90 - 96
[20] Crystal structure of the ATP-binding subunit of an ABC transporter
Hung, LW
Wang, IXY
Nikaido, K
Liu, PQ
Ames, GFL
Kim, SH
[J]. NATURE, 1998, 396 (6712) : 703 - 707

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