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 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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 131-136 |
| Number of pages | 6 |
| Journal | Nature biotechnology |
| Volume | 19 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 21 2001 |
Keywords
- Aggregation
- Complementation
- Folding
- Protein-folding diseases
- Solubility
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
- Molecular Medicine
- Biomedical Engineering