A new protamine-specific-metalloproteinase from Penicillium

一种来自青霉菌的新型鱼精蛋白特异性金属蛋白酶

• 批准号: 02660075
• 负责人: ICHISHIMA Eiji
• 金额: $ 1.34万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02660075/
• 关键词: Metalloproteinase; Neutral Proteinase; Penicillium; Protase; Specificity

It was found that a metalloproteinase from Penicillium citrinum has a specificity unique from those of other metalloproteinases on nuclear basic proteins and oxidized insulin B-chain.The enzyme was found to contain 1 gram-atom zinc per mole of enzyme with the molecular weight of 17, 000. About 95% of the amino acid sequence of the enzyme was determined. The circular dichroism(CD)of the holo- and apo-enzymes has been investigated. One gram-atom of zinc was essential component of the enzyme not only the activity but also the conformation.The enzyme was specifically active on basic nuclear protein& such as clupeine, salmine and histone at pH 7.0. The initial site of cleavage on the oxidized insulin B-chain by the proteinase was between Tyrl6-Leul7, and additional sites, Glul3-Alal4 and Alal4-Leul5 were noted. Hydrolyses of small peptides consisting 5 - 13 amino acids such as bradykinin, dynorphin-A, alpha-neoendorphin, neurotensin, luteinizing hormone releasing hormone, alpha-melanocyt. e stimulating hormone, substance P and chicken brain pentapeptide were noted.

结果表明，桔青霉的一种金属蛋白酶在核碱性蛋白和氧化胰岛素B链上具有不同于其他金属蛋白酶的特异性，每摩尔酶含有1克原子锌，相对分子质量为17000。测定了该酶约95%的氨基酸序列。研究了全酶和脱辅酶的圆二色谱(CD)。1克原子的锌是该酶活性和构象的必需组分，该酶在pH为7.0时对碱性核蛋白如马鞭草碱、盐碱和组蛋白具有特异活性。该酶在氧化的胰岛素B链上的起始切割位置在Tyr16-Leul7之间，另外还发现了Glul3-Alal4和Alal4-Leul5。由5-13个氨基酸组成的小肽，如缓激肽、强啡肽-A、α-新内啡肽、神经降压素、黄体生成素释放激素、α-黑素细胞的水解物。观察E刺激素、P物质和鸡脑五肽的变化。

项目成果

M.Kobayashi,M.Miura,T.Watanabe&E.Ichishima: "Affinity labeling of a subsite of Takaーamylase A by the fluorescent reagent Oーphthalaldehyde." Arch.Biochem.Biophys.289. 350-354 (1991)

M.Kobayashi、M.Miura、T.Watanabe&E.Ichishima：“荧光试剂邻苯二甲醛对 Taka-淀粉酶 A 亚位点的亲和标记。”Arch.Biochem.Biophys.289（1991）。

Y.Chiba,M.Ui,Y.Kato,T.Nakajima & E.Ichishima: "Two groups of intracellular αーamylase isoenzymes from cultured rice cells." Phytochemistry. 29. 2075-2078 (1990)

Y.Chiba、M.Ui、Y.Kato、T.Nakajima 和 E.Ichishima：“来自培养水稻细胞的两组细胞内 α-淀粉酶同工酶。” 29. 2075-2078 (1990)。

Y.Chiba,Y.Nieda,T.Nakajima & E.Ichishima: "Unique enzymatic properties of αーamylase III from suspension cultured rice cells." Agric.Biol.Chem.55. 901-902 (1991)

Y.Chiba、Y.Nieda、T.Nakajima 和 E.Ichishima：“悬浮培养的水稻细胞中 α-淀粉酶 III 的独特酶学特性。”Agric.Biol.Chem.55（1991）。

E. Ichishima: "Mode of action and application of Aspergillus carboxypeptidase." Comment Agric. & Food Chem.2. 279-298 (1991)

E. Ichishima：“曲霉羧肽酶的作用方式和应用。”

M. Kobayashi, M. Miura, T. Watanabe & E. Ichishima: "Affinity labeling of a subsite of Take-amylase A by the fluorescent reagent O-phthalaldehyde." Arch. Biochem. Biophys.289. 350-354 (1991)

M.小林、M.三浦、T.渡边