Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 177-182 |
| Number of pages | 6 |
| Journal | Biological chemistry |
| Volume | 386 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 1 2005 |
| Externally published | Yes |
Fingerprint
All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
- Clinical Biochemistry
Cite this
Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda : Comparative studies with blarina toxin. / Kita, Masaki; Okumura, Yuushi; Ohdachi, Satoshi D.; Oba, Yuichi; Yoshikuni, Michiyasu; Nakamura, Yasuo; Kido, Hiroshi; Uemura, Daisuke.
In: Biological chemistry, Vol. 386, No. 2, 01.02.2005, p. 177-182.Research output: Contribution to journal › Article
}
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
PY - 2005/2/1
Y1 - 2005/2/1
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.
UR - http://www.scopus.com/inward/record.url?scp=14944365562&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=14944365562&partnerID=8YFLogxK
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 -