title: Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda: comparative studies with blarina toxin.
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 (K(i) =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.
authors.affiliation: Research Center for Materials Science, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8602, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Kita
authors.forename: Yuushi
authors.initials: Y
authors.lastname: Okumura
authors.forename: Satoshi D
authors.initials: SD
authors.lastname: Ohdachi
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Oba
authors.forename: Michiyasu
authors.initials: M
authors.lastname: Yoshikuni
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kido
authors.forename: Daisuke
authors.initials: D
authors.lastname: Uemura
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D006863:Hydrogen-Ion Concentration; D007610:Kallikreins / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000494:pharmacology; D016415:Sequence Alignment; D012788:Shrews; D013379:Substrate Specificity; D020840:Tissue Kallikreins / Q000737:chemistry*
chemical_list: D011480:Protease Inhibitors; D007610:Kallikreins; C502041:blarinasin protein, Blarina brevicauda; D020840:Tissue Kallikreins; C487512:blarina toxin, Blarina brevicauda