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

• Published in: Nature biotechnology 2001-02, Vol.19 (2), p.131-136
• Main Authors: Thomas, Philip J, Wigley, W. Christian, Stidham, Rhesa D, Smith, Nathan M, Hunt, John F
• Format: Article
• Language: English
• Subjects: Agriculture; Amyloid beta-Peptides - chemistry; Amyloid beta-Peptides - genetics; Analytical, structural and metabolic biochemistry; beta-Galactosidase - chemistry; beta-Galactosidase - genetics; beta-Galactosidase - metabolism; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Cystic Fibrosis Transmembrane Conductance Regulator - chemistry; Cystic Fibrosis Transmembrane Conductance Regulator - genetics; E coli; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Genes, Reporter; Genetic engineering; Genetic Markers; Genetic technics; Humans; Life Sciences; Methods. Procedures. Technologies; Miscellaneous; Mutagenesis, Site-Directed; Peptide Fragments - chemistry; Protein Folding; Proteins; Publishing; Recombinant Fusion Proteins - chemistry; Solubility; Synthetic digonucleotides and genes. Sequencing
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/84389

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 α- and ω- fragments of β-galactosidase (β-gal). Fusions of the α-fragment to the C terminus of target proteins with widely varying in vivo folding yield and/or solubility levels, including the Alzheimer's amyloid β (Aβ) peptide and a non-amyloidogenic mutant thereof, reveal an unambiguous correlation between β-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.