Characteristics of monkey tryptase purified from cheek pouch vascular tissues.
Tryptase purified from rat and dog tissues has been reported, although the characteristics of these enzymes are different from human tryptase. For pathophysiological studies of human tryptase, studies on species that have a similar tryptase to humans is needed. In this study, we purified monkey tryptase from cheek pouch vascular tissues using heparin affinity and gel filtration columns. The monkey tryptase, which had a molecular weight of 130 kDa by gel filtration, consisted of a tetramer of 33 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The N-terminal sequence showed high homology with tryptases from other species. The optimum pH and temperature were 7.5-9.0 and 25-40 degrees C, respectively. The enzyme was labile in high-KCl buffer, and the optimum KCl concentration was 0.1 M. The enzyme activity was completely inhibited by diisopropyl phosphorofluoridate and leupeptin but not by soybean trypsin inhibitor and alpha-antitrypsin. The enzyme hydrolyzed vasoactive intestinal peptide but did not affect angiotensin I, somatostatin and bradykinin. In the present study, we first isolated monkey tryptase from cheek pouch vascular tissues and showed that the characteristics of monkey tryptase are very similar to those of human tryptase.
Sakaguchi
M
M
Department of Pharmacology, Osaka Medical College, Takatsuki City, Japan.
Takai
S
S
Jin
D
D
Yamada
M
M
Miyazaki
M
M
D000818:Animals; D002610:Cheek / Q000098:blood supply; D004795:Enzyme Stability; D008252:Macaca fascicularis; D008297:Male; D011189:Potassium Chloride / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000187:drug effects / Q000302:isolation & purification; D013045:Species Specificity; D014361:Trypsin Inhibitors / Q000494:pharmacology; D053802:Tryptases
D014361:Trypsin Inhibitors; D011189:Potassium Chloride; D012697:Serine Endopeptidases; C509223:Tpsab1 protein, rat; C509221:Tpsb2 protein, rat; D053802:Tryptases