Nature Biotechnology
19, 131 - 136 (2001)
doi:10.1038/84389
Protein solubility and folding monitored in vivo by structural complementation of a genetic marker proteinW. Christian Wigley1, Rhesa D. Stidham1, 2, Nathan M. Smith1, John F. Hunt3
& Philip J. Thomas1, 21
Department of Physiology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas TX 75235. 2
Graduate Program in Molecular Biophysics, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas TX 75235. 3
Department of Biological Sciences, Columbia University, New York NY 10027.
Correspondence should be addressed to Philip J. Thomas thomas07@utsw.swmed.eduaggregationfoldingsolubilitycomplementationprotein-folding diseasesProtein 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  - and  - fragments of  -galactosidase (-gal). Fusions of the -fragment to the C terminus of target proteins with widely varying in vivo folding yield and/or solubility levels, including the Alzheimer's amyloid (A) peptide and a non-amyloidogenic mutant thereof, reveal an unambiguous correlation between -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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