High-resolution X-ray crystal structures of human gammaD crystallin (1.25 A) and the R58H mutant (1.15 A) associated with aculeiform cataract.
Several human cataracts have been linked to mutations in the gamma crystallin gene. One of these is the aculeiform cataract, which is caused by an R58H mutation in gammaD crystallin. We have shown previously that this cataract is caused by crystallization of the mutant protein, which is an order of magnitude less soluble than the wild-type. Here, we report the very high-resolution crystal structures of the mutant and wild-type proteins. Both proteins crystallize in the same space group and lattice. Thus, a strict comparison of the protein-protein and protein-water intermolecular interactions in the two crystal lattices is possible. Overall, the differences between the mutant and wild-type structures are small. At position 58, the mutant protein loses the direct ion-pair intermolecular interaction present in the wild-type, due to the differences between histidine and arginine at the atomic level; the interaction in the mutant is mediated by water molecules. Away from the mutation site, the mutant and wild-type lattice structures differ in the identity of side-chains that occupy alternate conformations. Since the interactions in the crystal phase are very similar for the two proteins, we conclude that the reduction in the solubility of the mutant is mainly due to the effect of the R58H mutation in the solution phase. The results presented here are also important as they are the first high-resolution X-ray structures of human gamma crystallins.
Department of Crystallography, Birkbeck College, Malet Street, London WC1E 7HX, UK.
Ajit
A
Basak
Orval
O
Bateman
Christine
C
Slingsby
Ajay
A
Pande
Neer
N
Asherie
Olutayo
O
Ogun
George B
GB
Benedek
Jayanti
J
Pande
D019943:Amino Acid Substitution; D002386:Cataract / Q000235:genetics* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D006801:Humans; D066298:In Vitro Techniques; D008958:Models, Molecular; D009154:Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012995:Solubility; D055672:Static Electricity; D014867:Water / Q000737:chemistry; D038222:gamma-Crystallins / Q000737:chemistry* / Q000235:genetics*
D011994:Recombinant Proteins; D038222:gamma-Crystallins; D014867:Water