Differentiating human phospholipase A2's activity toward phosphatidylinositol, phosphatidylinositol phosphate and phosphatidylinositol bisphosphate

IF 3.9 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Daiki Hayashi , Edward A. Dennis
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引用次数: 0

Abstract

Phospholipase A2's (PLA2's) constitute a superfamily of enzymes that hydrolyze the sn-2 fatty acyl chain on glycerophospholipids. We have previously reported that each PLA2 Type shows a unique substrate specificity for the molecular species it hydrolyzes, especially the acyl chain that is cleaved from the sn-2 position and to some extent the polar group. However, phosphatidylinositol (PI) and PI phosphates (PIPs) have not been as well studied as substrates as other phospholipids because the PIPs require adaptation of the standard analysis methods, but they are important in vivo. We determined the in vitro activity of the three major types of human PLA2's, namely the cytosolic (c), calcium-independent (i), and secreted (s) PLA2's toward PI, PI-4-phosphate (PI(4)P), and PI-4,5-bisphosphate (PI(4,5)P2). The in vitro assay revealed that Group IVA cPLA2 (GIVA cPLA2) showed relatively high activity toward PI and PI(4)P among the tested PLA2's; nevertheless, the highly hydrophilic headgroup disrupted the interaction between the lipid surface and the enzyme. GIVA cPLA2 and GVIA iPLA2 showed detectable activity toward PI(4,5)P2, but it appeared to be a poorer substrate for all of the PLA2's tested. Furthermore, molecular dynamics (MD) simulations demonstrated that Thr416 and Glu418 of GIVA cPLA2 contribute significantly to accommodating the hydrophilic head groups of PI and PI(4)P, which could explain some selectivity for PI and PI(4)P. These results indicated that GIVA cPLA2 can accommodate PI and PI(4)P in its active site and hydrolyze them, suggesting that the GIVA cPLA2 may best account for the PI and PIP hydrolysis in living cells.

区分人类磷脂酶 A2 对磷脂酰肌醇、磷脂酰肌醇磷酸酯和磷脂酰肌醇二磷酸酯的活性。
磷脂酶 A2(PLA2)是水解甘油磷脂上 sn-2 脂肪酰基链的酶超家族。我们以前曾报道过,每种 PLA2 类型对其水解的分子种类都显示出独特的底物特异性,尤其是从 sn-2 位置裂解的酰基链,以及在一定程度上裂解的极性基团。然而,磷脂酰肌醇(PI)和磷脂酰肌醇磷酸酯(PIPs)作为底物的研究并不像其他磷脂那样深入,因为 PIPs 需要调整标准分析方法,但它们在体内却很重要。我们测定了人类 PLA2 的三种主要类型,即细胞质(c)、钙离子依赖型(i)和分泌型(s)PLA2 对 PI、PI-4-磷酸(PI(4)P)和 PI-4,5-二磷酸(PI(4,5)P2)的体外活性。体外试验显示,在所测试的 PLA2 中,第 IVA 组 cPLA2(GIVA cPLA2)对 PI 和 PI(4)P 的活性相对较高;然而,高亲水性头基破坏了脂质表面与酶之间的相互作用。GIVA cPLA2 和 GVIA iPLA2 对 PI(4,5)P2 具有可检测到的活性,但它似乎是所有受测 PLA2 的较差底物。此外,分子动力学(MD)模拟表明,GIVA cPLA2 的 Thr416 和 Glu418 在容纳 PI 和 PI(4)P 的亲水性头基方面起着重要作用,这可以解释对 PI 和 PI(4)P 的某些选择性。这些结果表明,GIVA cPLA2 能在其活性位点容纳 PI 和 PI(4)P 并水解它们,这表明 GIVA cPLA2 可能是活细胞中 PI 和 PIP 水解的最佳解释。
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来源期刊
CiteScore
11.00
自引率
2.10%
发文量
109
审稿时长
53 days
期刊介绍: BBA Molecular and Cell Biology of Lipids publishes papers on original research dealing with novel aspects of molecular genetics related to the lipidome, the biosynthesis of lipids, the role of lipids in cells and whole organisms, the regulation of lipid metabolism and function, and lipidomics in all organisms. Manuscripts should significantly advance the understanding of the molecular mechanisms underlying biological processes in which lipids are involved. Papers detailing novel methodology must report significant biochemical, molecular, or functional insight in the area of lipids.
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