Structural analysis for enzymatic activity of sphingomyelinase from Bacillus cereus

蜡状芽孢杆菌鞘磷脂酶酶活性的结构分析

• 批准号: 18590441
• 负责人: SAKURAI J.
• 金额: $ 2.57万
• 依托单位: Tokushima Bunri University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590441/
• 关键词: Bacillus cereus; sphingomyelinase; hemolytic activity; structural analysis; divalent cation; cleft; 3D structure; neutral SMase; セレウス菌; スフィンゴミエリン; アミノ酸置換

Neutral sphingomyelinase (nSMase) has been implicated in the generation of second messengers in mammalian cells. Bacillus cereus Smase (Bc-SMase) mimics the action of the endogenous mammalian nSMase in causing differentiation, development aging and apoptosis. The enzymatic activity of Bc-SMase was activated in the order of Co^<2+> > Mg^<2+> >> Ca^<2+>. The crystal structures of Bc-Smase with these metal ions were determined. The water-bridged twin divalent metal ions, Co^<2+> and Mg^<2+>, at the center of cleft were concluded to be the catalytic architecture to exert the sphingomyelinase activity. On the other hands, the architecture of Ca^<2+> binding at the site was different from that of Ca^<2+> and Mg^<2+>. The other binding site of these metal ions was at one side edge of the cleft. The crystal structure of the enzyme with Mg^<2+> or Co^<2+> at the site provides the universal structure bases for the hydrolytic activity of nSMase. In addition, the structural features and site-directed mutagenesis suggest that the specific-hairpin with the aromatic amino acid residues, the new active site, involves in binding to SM and membrane SM.

中性鞘磷脂酶（nSMase）参与哺乳动物细胞中第二信使的产生。蜡样芽孢杆菌Smase（Bc-SMase）模拟内源性哺乳动物nSMase在引起分化、发育、衰老和凋亡中的作用。Co^<2+> > Mg^<2+>> Ca ^<2+>对Bc-SMase酶活性的激活作用顺序为Co^<2+> > Mg^<2+>> Ca^<2+>。测定了Bc-Smase的晶体结构。位于裂隙中心的水桥双金属离子Co^2+和Mg^2+是发挥鞘磷脂酶活性的催化结构。另一方面，Ca^<2+>在该位点的结合结构不同于Ca^<2+>和Mg^<2+>。这些金属离子的另一个结合位点位于裂缝的一侧边缘。该酶的晶体结构中含有Mg^<2+>或Co^<2+>，为nSMase的水解活性提供了普遍的结构基础。此外，结构特征和定点突变表明，具有芳香族氨基酸残基的特异性发夹，新的活性位点，参与SM和膜SM的结合。

项目成果

The relationship between the metabolism of sphingomyelin species and hemolysis of sheep erythrocytes in duced by Clostridium perfringen alpha-toxin

鞘磷脂类代谢与产气荚膜梭菌α毒素引起的羊红细胞溶血的关系

• 发表时间: 2008
• 期刊: J.Lipid Res (In press)
• 作者: M.Oda;T.Matsuno;R.Shiihara;S.Ochi;R.Yamauchi;Y.Saito;H.Imagawa;M.Nagahama;M.Nishizawa and J.Sakural
• 通讯作者: M.Nishizawa and J.Sakural

セレウス菌スフィンゴミエリナーゼの細胞膜結合領域の解析

蜡样芽孢杆菌鞘磷脂酶细胞膜结合区的分析

• 发表时间: 2007
• 作者: 橋本 学;小田 真隆;津下 英明;勝沼 信彦;永浜 政博;櫻井 純
• 通讯作者: 櫻井 純

Structural basis of the sphingomyelin phosphodiesterase activity in neutral sphingomyelinase from Bacillus cereus

• DOI: 10.1074/jbc.m601089200
• 发表时间: 2006-06-09
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Ago, Hideo;Oda, Masataka;Sakurai, Jun
• 通讯作者: Sakurai, Jun

Clostridium perfringens beta-toxin; characterization and action

产气荚膜梭菌β-毒素；

• 发表时间: 2006
• 期刊: J.Toxicol.Toxin Rev. 25
• 作者: J.Sakurai;M.Nagahama
• 通讯作者: M.Nagahama

Oligomerization of Clostridium perfringens ε-toxin is dependent upon membrane fluidity in liposomes

• DOI: 10.1021/bi051805s
• 发表时间: 2006-01-10
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Nagahama, M;Hara, H;Sakurai, J
• 通讯作者: Sakurai, J