Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda: Comparative studies with blarina toxin

Masaki Kita, Yuushi Okumura, Satoshi D. Ohdachi, Yuichi Oba, Michiyasu Yoshikuni, Yasuo Nakamura, Hiroshi Kido, Daisuke Uemura

Research output: Contribution to journalArticle

24 Citations (Scopus)

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 languageEnglish
Pages (from-to)177-182
Number of pages6
JournalBiological chemistry
Volume386
Issue number2
DOIs
Publication statusPublished - Feb 1 2005
Externally publishedYes

Fingerprint

Shrews
Tissue Kallikreins
Kallikreins
Purification
Peptide Hydrolases
Salivary Glands
High Molecular Weight Kininogens
Aprotinin
Poisons
Venoms
Enzyme activity
Serine Proteases
Bradykinin
Enzymes
Amides
Complementary DNA
Amino Acids
Peptides

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 journalArticle

Kita, Masaki ; Okumura, Yuushi ; Ohdachi, Satoshi D. ; Oba, Yuichi ; Yoshikuni, Michiyasu ; Nakamura, Yasuo ; Kido, Hiroshi ; Uemura, Daisuke. / Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda : Comparative studies with blarina toxin. In: Biological chemistry. 2005 ; Vol. 386, No. 2. pp. 177-182.
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