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
[1]   HYALINIZATION OF THE ISLETS OF LANGERHANS IN DIABETES MELLITUS [J].
BELL, ET .
DIABETES, 1952, 1 (05) :341-344
[2]   Folding of a mutant maltose-binding protein of Escherichia coli which forms inclusion bodies [J].
Betton, JM ;
Hofnung, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (14) :8046-8052
[3]   A NOVEL STRATEGY FOR PRODUCTION OF A HIGHLY EXPRESSED RECOMBINANT PROTEIN IN AN ACTIVE FORM [J].
BLACKWELL, JR ;
HORGAN, R .
FEBS LETTERS, 1991, 295 (1-3) :10-12
[4]   Glycine betaine-assisted protein folding in a lysA mutant of Escherichia coli [J].
Bourot, S ;
Sire, O ;
Trautwetter, A ;
Touzé, T ;
Wu, LF ;
Blanco, C ;
Bernard, T .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (02) :1050-1056
[5]   Correcting temperature-sensitive protein folding defects [J].
Brown, CR ;
HongBrown, LQ ;
Welch, WJ .
JOURNAL OF CLINICAL INVESTIGATION, 1997, 99 (06) :1432-1444
[6]   Aggregation and motor neuron toxicity of an ALS-linked SOD1 mutant independent from wild-type SOD1 [J].
Bruijn, LI ;
Houseweart, MK ;
Kato, S ;
Anderson, KL ;
Anderson, SD ;
Ohama, E ;
Reaume, AG ;
Scott, RW ;
Cleveland, DW .
SCIENCE, 1998, 281 (5384) :1851-1854
[7]   PARTIAL DENATURATION OF TRANSTHYRETIN IS SUFFICIENT FOR AMYLOID FIBRIL FORMATION INVITRO [J].
COLON, W ;
KELLY, JW .
BIOCHEMISTRY, 1992, 31 (36) :8654-8660
[8]   Subcellular localization of the Alzheimer's disease amyloid precursor protein and derived polypeptides expressed in a recombinant yeast system [J].
Culvenor, JG ;
Henry, A ;
Hartmann, T ;
Evin, G ;
Galatis, D ;
Friedhuber, A ;
Jayasena, ULHR ;
Underwood, JR ;
Beyreuther, K ;
Masters, CL ;
Cappai, R .
AMYLOID-JOURNAL OF PROTEIN FOLDING DISORDERS, 1998, 5 (02) :79-89
[9]   Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation [J].
Davies, SW ;
Turmaine, M ;
Cozens, BA ;
DiFiglia, M ;
Sharp, AH ;
Ross, CA ;
Scherzinger, E ;
Wanker, EE ;
Mangiarini, L ;
Bates, GP .
CELL, 1997, 90 (03) :537-548
[10]   Protein misfolding, evolution and disease [J].
Dobson, CM .
TRENDS IN BIOCHEMICAL SCIENCES, 1999, 24 (09) :329-332
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