Kinetics of interfacial catalysis by phospholipase A2 in intravesicle scooting mode, and heterofusion of anionic and zwitterionic vesicles.

Kinetics of interfacial catalysis by phospholipase A2 in intravesicle scooting mode, and heterofusion of anionic and zwitterionic vesicles.
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磷脂酶 A2 在囊泡内移动模式下的界面催化动力学,以及阴离子和两性离子囊泡的异质融合。

DOI:
10.1016/0005-2736(86)90541-9
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发表时间:
1986
期刊:
Biochimica et biophysica acta
影响因子:
--
通讯作者:
Ramirez,F
Ramirez,F
中科院分区:
--
文献类型:
--
作者:
Jain,MK;Rogers,J;Jahagirdar,DV;Marecek,JF;Ramirez,F

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在这篇和下面的三篇论文中,我们研究了猪胰腺磷脂酶A2对无任何添加剂的阴离子磷脂囊泡的作用动力学。结果提供了第一个明确的证明,在囊泡内scooting模式的界面催化。在本文中,我们描述的条件下,猪胰腺磷脂酶A2对DMPMe(酯)囊泡的行动在没有任何添加剂的情况下开始没有潜伏期。在这些条件下,游离单体底物浓度是微不足道的;双层封闭的囊泡组织保持完整,即使当所有的基板在外层单层已被水解;囊泡间交换的速率和分子的transbilayer运动(触发器)的速率是可以忽略不计的慢;和囊泡的融合率是微不足道的。因此,与一个囊泡结合的酶分子通过一级过程水解囊泡外单层中的所有DMPMe分子,速率常数为30°C下每分钟0.6;或者从另一个角度来看,DMPMe囊泡中的一个酶分子可以以每分钟3000个的速率水解所有可用的底物分子。在低阴离子浓度下,过量的底物囊泡不会被水解,除非结合酶的囊泡间交换速率受水相中阴离子的刺激。较高的钙浓度不仅促进DMPMe囊泡的同源融合,而且促进DMPMe和DMPC囊泡的异源融合。有人提出,钙诱导的等温横向相分离在DMPMe囊泡诱导缺陷的双层组织,和这样的缺陷是磷脂酶A2结合和异源融合与DMPC(酯)囊泡,没有这样的网站。
In this and the following three papers we examine the kinetics of action of pig pancreatic phospholipase A2on vesicles of anionic phospholipids without any additives. The results provide the first unequivocal demonstration of interfacial catalysis in intravesicle scooting mode. In this paper we describe the conditions in which the action of pig pancreatic phospholipase A2on DMPMe (ester) vesicles in the absence of any additive commences without a latency. Under these conditions the free monomer substrate concentration is insignificant; the bilayer enclosed vesicle organization remains intact even when all the substrate in the outer monolayer has been hydrolyzed; the rate of intervesicle exchange and the rate of transbilayer movement (flip-flop) of molecules is negligibly slow; and the rate of fusion of vesicles is insignificant. Thus an enzyme molecule bound to one vesicle hydrolyzes all the DMPMe molecules in the outer monolayer of the vesicle by a first-order process with a rate constant of 0.6 per min at 30°C; or viewed another way, one enzyme molecule in a DMPMe vesicle can hydrolyze all the available substrate molecules at the rate of 3000 per min. At low anion concentrations excess substrate vesicles are not hydrolyzed unless the rate of intervesicle exchange of the bound enzyme is stimulated by anions in the aqueous phase. Higher calcium concentrations promote not only homofusion of DMPMe vesicles but also heterofusion of DMPMe and DMPC vesicles. It is proposed that calcium-induced isothermal lateral phase separation in DMPMe vesicles induces defects in the bilayer organization, and such defects are the sites for phospholipase A2binding and for heterofusion with DMPC (ester) vesicles which do not have such sites.
脂质双层囊泡融合的反应特点及机制
DOI: --
发表时间: 1984
期刊:
影响因子: --
作者:
S. M. Gibson;G. Strauss
通讯作者: G. Strauss
磷脂酶 A2 对链烷醇修饰的磷脂酰胆碱作用期间的滞后期。
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发表时间: 1978
影响因子: 4.8
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从人血小板中分离均质磷脂酶 A2。
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发表时间: 1979
影响因子: 3.1
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R. Apitz;M. Mas;M. Cruz;M. Jain
通讯作者: M. Jain
DOI: --
发表时间: 1985
期刊: Biochemistry
影响因子: 2.9
作者:
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通讯作者: D. Zakim
磷脂酶 A2 的多相催化:表面效应动力学描述的公式化。
DOI: 10.1139/o79-012
发表时间: 1979
期刊: Canadian journal of biochemistry
影响因子: --
作者:
D. Tinker;J. Wei
通讯作者: J. Wei