CHARACTERIZATION OF 2 SERPINS FROM BOVINE PLASMA AND MILK

An inhibitor of pancreatic elastase (EI), which can also inhibit chymotrypsin, and an inhibitor of trypsin (TI), which can also inhibit plasmin, have been isolated from bovine plasma. EI and TI belong to the serpin family of inhibitors. The size of both inhibitors is approx. 60 kDa and they are able to form SDS-stable complexes with proteinases. Curiously, TI dimerizes in the presence of SDS, a feature which has been observed previously only in non-denaturing gels of human alpha(1)-antitrypsin (alpha(1)PI). EI and TI are glycosylated [16% and 19% (w/w) respectively] and their amino acid compositions are similar to those of other plasma serpins. Neither EI nor TI is the equivalent of bovine alpha(1)PI, as revealed by partial sequence analysis of their N-termini and reactive sites. Rather, both inhibitors appear to be related to human alpha(1)-antichymotrypsin. Inhibition of pancreatic elastase and chymotrypsin by EI occurs with a k(ass.) similar to 10(5) M(-1).s(-1). TI inhibits trypsin with a k(ass.) similar to 10(5) M(-1).s(-1). Plasmin is inhibited by TI with a k(ass.) 10(3) M(-1).s(-1). The values of the kinetic constants are similar to those determined for the well-studied human serpins. Antibodies to EI and TI reveal a set of four antigenically related proteins of similar size in plasma. In addition, they detect the same set of proteins in milk. The inhibitors isolated from milk are identical to EI and TI from plasma. EI could control the activity of chymotrypsin-like proteinases in milk. In contrast, no target proteinases of TI in milk can be suggested.