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
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
收藏
页码:131 / 136
页数:6
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