Molecular Theory of the Catalytic Function of Phospholipase A2

磷脂酶A2催化功能的分子理论

基本信息

  • 批准号:
    02671022
  • 负责人:
  • 金额:
    $ 1.66万
  • 依托单位:
  • 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 1992
  • 项目状态:
    已结题

项目摘要

1. Kinetics of the hydrolysis of monodispersed and micellar sustrates, catalyzed by Group I or II phospholipase A_2 (PLA_2s) in the presence of a saturating Ca^<2+> concentration indicated that deprotonated state of the catalytic group His 48 and protonated state of Tyr 52, the site of which is located in close proximity to the imidazole ring of His 48 were found to be essential for the catalysis. The importance of an ionized state of the N-terminal alpha-amino group at the active site was also indicated for Group I enzymes. The pK values of both His 48 and Tyr 52 of the enzyme-Ca^<2+> complex shifted markedly to the alkaline side on binding to micellar substrates, whereas no significant pK shifts were noted for the bindings to monodispersed substrates.2. Kinetic studies showed that Ca^<2+> binding to the both types of PLA_2s was essential for the catalysis. Substrate bindings to Group I PLA_2s were independent of the Ca^<2+> binding, whereas those for Group II enzymes were facilitated … More more than 10 times by the Ca^<2+> binging. The latter result was compatible with the hypothesis that an intermediate complex would be stabilized by coordination of the bound Ca^<2+> ion with the phosphoryl group and the carbonyl group at the sn-2 position of the substrate molecule. However, the former, the former result for Group I enzymes seemed incompatible for this mechanism. The X-ray crystallographic studies on the bindings of a substrate analog having an amide-bond instead of the sn-2 ester bond, dodecanoyl-2-aminohexanol-1-phosphoglycol, indicated that the carbonyl oxygen and phosphoryl moiety interact directly with the bound Ca^<2+> ion at the substrate binding site, suggesting that the binding of this analog should depend on the Ca^<2+> binding. Our studies in solution to confirm this showed the binding constants of this analog to both types of enzymes were increased significantly as the degrees of Ca^<2+> bindings increase, indicating that the structures of enzyme-substrate analog complex and enzyme-Ca^<2+>-analog complex in solution are very similar to each other as those in crystal and that the structures of the enzyme-analog complexes are stabilized by the Ca^<2+> binding. The enzyme-Ca^<2+>-genuine substrate complexes for the both types of PLA_2s are also considered to have similar structures.3. Some metal ions including lantanide ions were showed to bind to PLA_2s and to their genuine-substrate complexes in a manner similar to that of Ca^<2+> ion and to produce the PLA_2 activites (less than 25%). Less
1.在饱和Ca~(2+)浓度下,磷脂酶A_2催化单分散和胶束底物的水解动力学表明,催化基团His 48的去质子化状态和酪氨酸52的质子化状态是催化反应所必需的。对于I组酶,活性部位N-末端α-氨基的电离状态的重要性也被指出。在与胶束底物结合时,酶-Ca^&lt;2+&gt;络合物的His 48和Tyr 52的pK值均显著向碱性侧移动,而与单分散底物的结合没有明显的pk移动。动力学研究表明,Ca~(2+)和~(2+)与两种类型的磷脂酶A_2S的结合是催化反应所必需的。与第一组磷脂酶A_2S的底物结合不依赖于与钙离子的结合,而与第二组酶的底物结合则促进了…超过10倍的钙^&lt;2+;狂欢。后者的结果与以下假设是一致的,即通过结合的Ca^&lt;2+&gt;离子与底物分子sn-2位的磷酰基和羰基配位来稳定中间络合物。然而,第一组酶的前一种结果似乎与这一机制不相容。对具有酰胺键而不是sn-2酯键的底物类似物的结合进行的X-射线结晶学研究表明,羰基氧和磷酰基部分直接与底物结合部位的结合钙离子相互作用,表明该类似物的结合应依赖于钙离子的结合。我们在溶液中的研究证实了这一点,该类似物对这两种酶的结合常数随着Ca^&lt;2+&gt;结合度的增加而显著增加,表明酶-底物类似物复合体和酶-类似物复合体在溶液中的结构与晶体中的结构非常相似,酶-类似物复合体的结构是由Ca^&lt;2+&gt;结合所稳定的。这两种类型的PLA2的酶-Ca^&lt;2+&gt;-真底物络合物也被认为具有相似的结构。一些金属离子,包括Lantanide离子,以类似于Ca~(2+)~(2+)和Gt~(2+)离子的方式与PLA2及其真正底物络合物结合,并产生PLA2活性(低于25%)。较少

项目成果

期刊论文数量(12)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Shuji Hada: "Hydrolysis of Micellar Diheptanoylphosphatidylcholine Catalyzed by Bovine Pancreatic Phospholipase A_2:Kinetic Characterization of Group I and II Enzymes" J.Biochem.113. 13-18 (1993)
Shuji Hada:“牛胰磷脂酶 A_2 催化胶束二庚酰磷脂酰胆碱的水解:I 组和 II 组酶的动力学表征”J.Biochem.113。
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    0
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KEIZO TESHIMA: "Kinetics of the Hydrolysis of Mixed Micelles of Dipalmitoyllecithin with Triton Xー100 Catalyzed by a Phospholipase A_2 from the Venom of Agkistrodon halys blomhoffii" Journal of Biochemistry. 108. 21-27 (1990)
KEIZO TESHIMA:“来自 Agkistrodon halys blomhoffii 毒液的磷脂酶 A_2 催化 Triton X-100 水解二棕榈酰卵磷脂混合胶束的动力学”《生物化学杂志》108. 21-27 (1990)。
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  • 发表时间:
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  • 影响因子:
    0
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Shuji HADA: "Hydrolysis of Micellar Ditheptanoylphosharidylcholine Catalyzed by Bovine Pancreatic Phospholipase A_2:Kinetic Characterization of Group I and II Enzymes" J.Biochem.113. 13-18 (1993)
Shuji HADA:“牛胰磷脂酶 A_2 催化胶束二特庚酰磷脂酰胆碱的水解:I 组和 II 组酶的动力学表征”J.Biochem.113。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
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Shuji Hada: "Hydrolysis of Micellar Diheptanoylphosphatidylcholine Catalyzed by Bovine Pancreatic Phospholipase A_2 : Kinetic Characterization of Group I and II Enzymes" J.Biochem. 113(1). 13-18 (1993)
Shuji Hada:“牛胰磷脂酶 A_2 催化胶束二庚酰磷脂酰胆碱的水解:I 组和 II 组酶的动力学表征”J.Biochem。
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  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Shinobu FUJII: "Kinetics of the Hydrolysis of Monodispersed Dihexanoylphosphatidylcholine Catalyzed by Bovine Pancreatic Phospholipase A_2:Roles of Ca^<2+> Binding and Ionizations of Amino Acid Residues in the Active Site" J.Biochem.110. 1008-1015 (1991)
Shinobu FUJII:“牛胰磷脂酶 A_2 催化的单分散二己酰磷脂酰胆碱的水解动力学:活性位点中 Ca^2 > 结合和氨基酸残基电离的作用”J.Biochem.110。
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    0
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IKEDA Kiyoshi其他文献

IKEDA Kiyoshi的其他文献

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{{ truncateString('IKEDA Kiyoshi', 18)}}的其他基金

Synthesis and Biological Evaluations as Inhibitors of Human Parainfluenza Virus-1 based on the Computational Design for Prevention of Multi-infectious Diseases
基于预防多种传染病的计算设计的人类副流感病毒1抑制剂的合成和生物学评价
  • 批准号:
    24590155
  • 财政年份:
    2012
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Sialic acid derivative synthesis and inhibitory activities against human parainfluenza virus type 1
唾液酸衍生物的合成及其对人副流感病毒1型的抑制活性
  • 批准号:
    21590117
  • 财政年份:
    2009
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Synthesis of sialic acid derivatives for human parainfluenza virus type-1 sialidase inhibitors
人副流感病毒1型唾液酸酶抑制剂唾液酸衍生物的合成
  • 批准号:
    19590103
  • 财政年份:
    2007
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Chemoenzymatic synthesis of N-acetylneuraminic acid derivatives and their behaviour towards sialidase from human parainfluenza virus
N-乙酰神经氨酸衍生物的化学酶合成及其对人副流感病毒唾液酸酶的行为
  • 批准号:
    13672223
  • 财政年份:
    2001
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Development of simple apparatus for measuring liquid-concentration by ultrasonic light diffraction effect
利用超声波光衍射效应测量液体浓度的简易装置的研制
  • 批准号:
    12650051
  • 财政年份:
    2000
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Elucidation of the Catalytic Function of Phospholipases
磷脂酶催化功能的阐明
  • 批准号:
    09672264
  • 财政年份:
    1997
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Synthesis of Tn and Sialyl Tn Antigen-Lipid A Analog Conjugate for Synthetic Vaccines
用于合成疫苗的 Tn 和唾液酸 Tn 抗原-脂质 A 类似物缀合物的合成
  • 批准号:
    08672565
  • 财政年份:
    1996
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Elucidation of the Catalytic Function of Phospholipase A_2 and C
磷脂酶 A_2 和 C 催化功能的阐明
  • 批准号:
    07672399
  • 财政年份:
    1995
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Elucidation of the Catalytic Function and the Inhibition mechanism of Phospholipase A_2
磷脂酶A_2的催化功能和抑制机制的阐明
  • 批准号:
    05671853
  • 财政年份:
    1993
  • 资助金额:
    $ 1.66万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)

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结直肠癌干细胞线粒体内钙离子动态靶向疗法的开发
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Evaluation of voltage-gated calcium ion channels as a therapeutic target in intrahepatic cholangiocarcinoma
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钙离子通道在胶原重塑中的作用
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合作研究:CRCNS 研究提案:控制 NMJ 的 AP 波形、钙离子、进入和递质释放的突触前结构功能关系
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