In a recent paper, patients with a progressive juvenile-onset hereditary ca
taract have been reported to have a point mutation in the human gamma D cry
stallin gene (Stephan, D. A., Gillanders, E., Vanderveen, D., Freas-Lutz, D
., Wistow, C., Baxevanis, A. D., Robbins, C. M., VanAuken, A., Quesenberry,
M. I., Bailey-Wilson, J., ef al. (1999) Proc. Natl. Acad. Sci. USA 96, 100
8-1012). This mutation results in the substitution of Arg-14 in the native
protein by a Cys residue. It is not understood how this mutation leads to c
ataract. We have expressed recombinant wild-type human gamma D crystallin (
HGD) and its Arg-14 to Cys mutant (R14C) in Escherichia coli and show that
R14C forms disulfide-linked oligomers, which markedly raise the phase separ
ation temperature of the protein solution. Eventually, R14C precipitates. I
n contrast, HGD slowly forms only disulfide-linked dimers and no oligomers.
These data strongly suggest that the observed cataract is triggered by the
thiol-mediated aggregation of R14C. The aggregation profiles of HGD and R1
4C are consistent with our homology modeling studies that reveal that R14C
contains two exposed cysteine residues, whereas HGD has only one. Our CD, f
luorescence, and differential scanning calorimetric studies show that HGD a
nd R14C have nearly identical secondary and tertiary structures and stabili
ties. Thus, contrary to current views, unfolding or destabilization of the
protein is not necessary for cataractogenesis.