TY - JOUR
T1 - Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda
T2 - Comparative studies with blarina toxin
AU - Kita, Masaki
AU - Okumura, Yuushi
AU - Ohdachi, Satoshi D.
AU - Oba, Yuichi
AU - Yoshikuni, Michiyasu
AU - Nakamura, Yasuo
AU - Kido, Hiroshi
AU - Uemura, Daisuke
N1 - Funding Information:
This research was supported in part by Grants-in-Aid for Scientific Research on Priority Area (A) and for Creative Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (to D.U.). The Blarina shrew project was conducted on property owned by the University of Michigan and in collaboration with Dr. P. Myers. We thank Dr. K. Suenaga and Dr. S. Kawada (Nagoya University), A.P.G. Dowling (University of Michigan), A. Uesugi and F. Okabe (Hokkaido University) for collecting Blarina specimens.
PY - 2005/2
Y1 - 2005/2
N2 - A new tissue kallikrein-like protease, blarinasin, has been purified from the salivary glands of the short-tailed shrew Blarina brevicauda. Blarinasin is a 32-kDa N-glycosylated protease with isoelectric values ranging between 5.3 and 5.7, and an optimum pH of 8.5 for enzyme activity. The cloned blarinasin cDNA coded for a pre-pro-sequence and a mature peptide of 252 amino acids with a catalytic triad typical for serine proteases and 43.7-54.0% identity to other mammalian tissue kallikreins. Blarinasin preferentially hydrolysed Pro-Phe-Arg-4-methylcoumaryl-7-amide (MCA) and N-tert-butyloxycarbonyl-Val-Leu-Lys-MCA, and preferentially converted human high-molecular-weight kininogen (HK) to bradykinin. The activity of blarinasin was prominently inhibited by aprotinin Ki=3.4 nM). A similar kallikrein-like protease, the lethal venom blarina toxin, has previously been purified from the salivary glands of the shrew Blarina and shows 67.9% identity to blarinasin. However, blarinasin was not toxic in mice. Blarinasin is a very abundant kallikrein-like protease and represents 70-75% of kallikrein-like enzymes in the salivary gland of B. brevicauda.
AB - A new tissue kallikrein-like protease, blarinasin, has been purified from the salivary glands of the short-tailed shrew Blarina brevicauda. Blarinasin is a 32-kDa N-glycosylated protease with isoelectric values ranging between 5.3 and 5.7, and an optimum pH of 8.5 for enzyme activity. The cloned blarinasin cDNA coded for a pre-pro-sequence and a mature peptide of 252 amino acids with a catalytic triad typical for serine proteases and 43.7-54.0% identity to other mammalian tissue kallikreins. Blarinasin preferentially hydrolysed Pro-Phe-Arg-4-methylcoumaryl-7-amide (MCA) and N-tert-butyloxycarbonyl-Val-Leu-Lys-MCA, and preferentially converted human high-molecular-weight kininogen (HK) to bradykinin. The activity of blarinasin was prominently inhibited by aprotinin Ki=3.4 nM). A similar kallikrein-like protease, the lethal venom blarina toxin, has previously been purified from the salivary glands of the shrew Blarina and shows 67.9% identity to blarinasin. However, blarinasin was not toxic in mice. Blarinasin is a very abundant kallikrein-like protease and represents 70-75% of kallikrein-like enzymes in the salivary gland of B. brevicauda.
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U2 - 10.1515/BC.2005.022
DO - 10.1515/BC.2005.022
M3 - Article
C2 - 15843162
AN - SCOPUS:14944365562
VL - 386
SP - 177
EP - 182
JO - Biological Chemistry
JF - Biological Chemistry
SN - 1431-6730
IS - 2
ER -