title: Site of carbamoylation of bovine gamma-II-crystallin by potassium [14C]cyanate.
abstract: One possible route to cataract formation may be via the carbamoylation of lens proteins due to increased concentrations of cyanate in the body resulting from uraemia associated with renal failure and with severe diarrhoea. Carbamoylation of gamma-II-crystallin, which is found in the lens core, could alter the surface charge network of the molecules, resulting in aggregation, increased light-scattering and hence cataract. We have attempted to locate the site(s) of carbamoylation in gamma-II-crystallin. gamma-II-Crystallin was isolated by gel chromatography and ion-exchange chromatography. gamma-II-Crystallin was then carbamoylated by incubation with potassium [14C]cyanate, followed by citraconylation and digestion with trypsin to give peptides that were separated by high-resolution ion-exchange chromatography. The amino acid compositions of the radioactive peptides were compared with the expected peptide composition for gamma-II-crystallin. The radioactive peptide compositions, which agreed with the theoretical peptides, all matched with the N-terminal region of gamma-II-crystallin and had in common the presence of the N-terminal glycine residue. It appears that the alpha-amino group of the N-terminal glycine was the main site of carbamoylation. This site forms part of the charge network on the surface of gamma-II-crystallin.
authors.affiliation: Nuffield Laboratory of Ophthalmology, University of Oxford, U.K.
authors.forename: S
authors.initials: S
authors.lastname: Martin
authors.forename: J J
authors.initials: JJ
authors.lastname: Harding
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002949:Citraconic Anhydrides; D003459:Crystallins / Q000378:metabolism*; D003485:Cyanates / Q000378:metabolism*; D010104:Oxygen Radioisotopes; D011485:Protein Binding; D011489:Protein Denaturation / Q000187:drug effects; D014357:Trypsin / Q000032:analysis
chemical_list: D000596:Amino Acids; D002949:Citraconic Anhydrides; D003459:Crystallins; D003485:Cyanates; D010104:Oxygen Radioisotopes; D014357:Trypsin; C024835:potassium cyanate