βB1 crystallin is an amine-donor substrate for tissue transglutaminase

The effects of tissue transglutaminase on the water-soluble proteins in bovine lens homogenates are described. Addition of liver transglutaminase and Ca²⁺ to calf lens homogenates resulted not only in the appearance of 50- and 57-kDa dimers, but also in a decrease in the amount of βB1 crystallin and the almost complete disappearance of βB3 and βA3. This is not the result of Ca²⁺-induced proteolysis, since histamine completely inhibits this phenomenon. It may be concluded that these polypeptides are involved in β-crystallin crosslinking by transglutaminase. This notion was confirmed by using βB1- and βBp-specific antisera. Both sera reacted with the 57-kDa dimer; the βBp-specific antiserum also reacted with the 50-kDa dimer. No reaction in the region 50–57 kDa was detectable when EDTA was used instead of Ca²⁺. Using reconstituted mixtures of βB1- and βBp-crystallin chains, and N-terminally truncated derivatives thereof, it was shown that in the βB1/βBp dimer, glutamine residue -9 of βBp crosslinks to one of the lysine residues in the N-terminal extension of βB1.