Home LiteratureArticle Details
PMID: 11175726 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Nature biotechnology ·Vol. 19 ·No. 2 ·2001-02-00 ·Pages 131-6

Wigley WC, Stidham RD, Smith NM, Hunt JF, Thomas PJ

Abstract

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.

MeSH Terms
Amyloid beta-Peptides/chemistry,genetics Cystic Fibrosis Transmembrane Conductance Regulator/chemistry,genetics Genes, Reporter Genetic Markers Humans Mutagenesis, Site-Directed Peptide Fragments/chemistry Protein Folding Recombinant Fusion Proteins/chemistry Solubility beta-Galactosidase/chemistry,genetics,metabolism
Chemicals
Amyloid beta-Peptides CFTR protein, human Genetic Markers Peptide Fragments Recombinant Fusion Proteins Cystic Fibrosis Transmembrane Conductance Regulator beta-Galactosidase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wigley W C
Department of Physiology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas TX 75235, USA.
Stidham R D
Smith N M
Hunt J F
Thomas P J
Article Info
Journal
Nature biotechnology
Abbr.
Nat Biotechnol
ISSN
1087-0156
Published
2001-02-00
Pages
131-6
Language
English
Region
United States
NLM ID
9604648
Subset
IM
Corrections
CommentIn
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com