Wc. Wigley et al., Protein solubility and folding monitored in vivo by structural complementation of a genetic marker protein, NAT BIOTECH, 19(2), 2001, pp. 131-136
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. T
he 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 b
eta (A beta) peptide and a non-amyloidogenic mutant thereof, reveal an unam
biguous correlation between beta -gal activity and the solubility/folding o
f the target. Thus, structural complementation provides a means of monitori
ng protein solubility/misfolding in vivo, and should find utility in the sc
reening for compounds that influence the pathological consequences of these
processes.