Activation of kallikrein-kinin system in human plasma with purified serine protease from Dermatophagoides farinae.
An aqueous extract from a mite culture, of Dermatophagoides farinae, activated prekallikrein to kallikrein in normal plasma. Crude protein preparation, obtained by ammonium sulfate precipitation (95% saturation) from the extract, exhibited high activity (0.81 units/mg protein) towards a synthetic substrate of coagulation factor XIIa, Boc-Gln-Gly-Arg-MCA, and had also activity to form kallikrein in human plasma deficient in coagulation factor XII. Treatment of the protein preparation with phenylmethylsulfonyl fluoride (PMSF), an inhibitor of serine enzyme, gave rise to inactivation of both activities. Thus, the serine protease specific for Boc-Gln-Gly-Arg-MCA in mite cultures of D. farinae was purified by ammonium sulfate precipitation and chromatographies on p-aminobenzamidine-sepharose CL-4B, DEAE-Toyopearl 650M, Sephadex G-75 superfine and Sephacryl S-200. The purified protease was homogeneous electrophoretically, and its molecular weight was estimated to be 30,000. The optimum pH and temperature were around 7.5 and 50 degrees C, respectively. The specific activity was 36 units/mg protein at pH 7.4 and 37 degrees C. The activity was completely inhibited by PMSF. The serine protease had the activity to activate the kallikrein-kinin system in normal human plasma.
Takahashi
K
K
Sagami Chemical Research Center, Juntendo University, School of Medicine, Japan.
Aoki
T
T
Kohmoto
S
S
Nishimura
H
H
Kodera
Y
Y
Matsushima
A
A
Inada
Y
Y
D000818:Animals; D004789:Enzyme Activation / Q000187:drug effects; D015956:Factor XIIa / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007610:Kallikreins / Q000097:blood*; D007700:Kinetics; D007705:Kinins / Q000097:blood*; D008925:Mites / Q000201:enzymology*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D013379:Substrate Specificity; D013696:Temperature
D007705:Kinins; D010664:Phenylmethylsulfonyl Fluoride; D007610:Kallikreins; D012697:Serine Endopeptidases; D015956:Factor XIIa