The quaternary structure of bovine alpha-crystallin. Chemical crosslinking with bifunctional imido esters.
Reaction of calf alpha-crystallin, which consists of about 30 A and 10 B subunits, with various bisimido esters at pH 8.0-8.5 and room temperature leads to inter-subunit crosslinking. Little difference is observed in the efficiency of crosslinking by reagents of a homologous series with different lengths (from C6 to C12). At any stage of the reaction an exponential decrease is found in the amounts of crosslinked dimer, trimer, tetramer etc., with no preference for the formation of any particular oligomer, suggesting that all subunits on the surface of the spherical alpha-crystallin molecule are in quasi-equivalent positions. Molecular weight analysis by docdecylsulfate gel electrophoresis and reversible crosslinking show that both A and B subunits occur in an apparently constant ratio in the crosslinked oligomers, from which we infer that both types of subunits are randomly arranged on the surface of the alpha-crystallin molecule. Not all of the subunits can form inter-chain crosslinks, as a limiting value of about 60% of the subunits is found in oligomeric form. A simple explanation could be a core of subunits which is inaccessible to certain chemical reagents, although other alternatives are also discussed.
D000818:Animals; D002417:Cattle; D003432:Cross-Linking Reagents; D003459:Crystallins; D004120:Dimethyl Suberimidate; D004591:Electrophoresis, Polyacrylamide Gel; D007096:Imidoesters; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011487:Protein Conformation; D013329:Structure-Activity Relationship
D003432:Cross-Linking Reagents; D003459:Crystallins; D007096:Imidoesters; D046911:Macromolecular Substances; D004120:Dimethyl Suberimidate