title: Protein solubility and folding monitored in vivo by structural complementation of a genetic marker protein.
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.
authors.affiliation: Department of Physiology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas TX 75235, USA.
authors.forename: W C
authors.initials: WC
authors.lastname: Wigley
authors.forename: R D
authors.initials: RD
authors.lastname: Stidham
authors.forename: N M
authors.initials: NM
authors.lastname: Smith
authors.forename: J F
authors.initials: JF
authors.lastname: Hunt
authors.forename: P J
authors.initials: PJ
authors.lastname: Thomas
mesh_terms: D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry* / Q000235:genetics; D017930:Genes, Reporter; D005819:Genetic Markers; D006801:Humans; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D012995:Solubility; D001616:beta-Galactosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; C505032:CFTR protein, human; D005819:Genetic Markers; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D001616:beta-Galactosidase