Masato Sakaguchi, Shinji Takai, Denan Jin, Mayumi Yamada and Mizuo Miyazaki*
Department of Pharmacology, Osaka Medical College, Takatsuki City, Osaka
569-8686, Japan
*Corresponding author.ﾊﾊFAX:+81-726-84-6518
E-mail: pha001@art.osaka-med.ac.jp
Abstract: 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ﾊkDa by gel filtration, consisted of a tetramer of 33ﾊ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｡C, respectively. The enzyme was labile
in high-KCl buffer, and the optimum KCl concentration was 0.1ﾊM. The enzyme
activity was completely inhibited by diisopropyl phosphorofluoridate and
leupeptin but not by soybean trypsin inhibitor and a-antitrypsin.
The enzyme hydrolyzed vasoactive intestinal peptide but did not affect angiotensinﾊ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.
Keywords: Tryptase, Monkey, Purification, Mast cell, Characteristic
Copyrightｩ The Japanese Pharmacological Society 2000
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