Angiotensin II generation by mast cell alpha- and beta-chymases.

Mast cells secrete alpha- and beta-chymases. Primate alpha-chymases generate angiotensin (AT) II by selectively hydrolyzing AT I's Phe(8)-His(9) bond. This is distinct from the AT converting enzyme (ACE) pathway. In humans, alpha-chymase is the major non-ACE AT II-generator. In rats, beta-chymases destroy AT II by cleaving at Tyr(4)-Ile(5). Past studies predicted that AT II production versus destruction discriminates alpha- from beta-chymases and that Lys(40) in the substrate-binding pocket determines alpha-chymase Phe(8) specificity. This study examines these hypotheses by comparing AT II generation by human alpha-chymase (containing Lys(40)), dog alpha-chymase (lacking Lys(40)), and mouse mMCP-4 (a beta-chymase lacking Lys(40); orthologous to AT II-destroying rat chymase rMCP-1). The results suggest that human and dog alpha-chymase generate AT II exclusively and with comparable efficiency, although dog chymase contains Ala(40) rather than Lys(40). Furthermore, AT II is the major product generated by degranulation supernatants from cultured dog mast cells, which release tryptases and dipeptidylpeptidase as well as alpha-chymase. In contrast to rMCP-1, mMCP-4 beta-chymase readily generates AT II. Although there is competing AT I hydrolysis at Tyr(4), mMCP-4 does not destroy AT II quickly once it is formed. We conclude (1) that chymases are the dominant AT I-hydrolyzing mast cell peptidases, (2) that residues other than Lys(40) are key determinants of alpha-chymase AT I Phe(8) specificity, (3) that beta-chymases can generate AT II, and (4) that alpha- and beta-chymases are not strictly dichotomous regarding AT I cleavage specificity.