Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda:: comparative studies with blarina toxin
Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda:: comparative studies with blarina toxin
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DOI：
10.1515/bc.2005.022
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发表时间：
2005-02-01
影响因子：
3.7
通讯作者：
Uemura, D
中科院分区：
文献类型：
--
作者：
Kita, M;Okumura, Y;Uemura, D
一种新的组织激肽释放酶样蛋白酶，blarinasin，是从短尾鼩鼱 Blarina brevicauda 的唾液腺中纯化出来的。 Blarinasin 是一种 32 kDa N-糖基化蛋白酶，等电值范围在 5.3 至 5.7 之间，酶活性的最佳 pH 值为 8.5。克隆的 blarinasin cDNA 编码前原序列和 252 个氨基酸的成熟肽，具有丝氨酸蛋白酶典型的催化三联体，与其他哺乳动物组织激肽释放酶具有 43.7-54.0% 的同一性。 Blarinasin 优先水解 Pro-Phe-Arg-4-methylcoumaryl-7amide (MCA) 和 N-叔丁氧基羰基-Val-Leu-Lys-MCA，并优先将人高分子量激肽原 (HK) 转化为缓激肽。 blarinasin 的活性被抑肽酶显着抑制 (K-i=3.4 nm)。一种类似的激肽释放酶样蛋白酶，即致命毒液 blarina 毒素，之前已从鼩鼱 Blarina 的唾液腺中纯化出来，与 blarinasin 具有 67.9% 的同一性。然而，blarinasin 对小鼠没有毒性。 Blarinasin 是一种非常丰富的激肽释放酶样蛋白酶，占短尾拟杆菌唾液腺中激肽释放酶样酶的 70-75%。
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-7amide (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.