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The ABC's of Group IV cytosolic phospholipase A_2

机译:IV组胞质磷脂酶A_2的ABC

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摘要

The three known human Group IV phospholipase A_2 (PLA_2) paralogs, Group IVA, 1VB and IVC, were overexpressed in Sf9 insect cells using the baculovirus expression system. An endogenous, calcium-independent PLA_2 activity was identified in the insect cell lysates, which can be inhibited by bromoenol lactone (BEL). The Group IV PLA_2 enzymes were characterized in overexpressing insect cell lysates in the presence of BEL, enabling their differentiation from the endogenous PLA_2 activity. Group IVC PLA_2 was found to have significant lysophospholipase activity, while Group IVB PLA_2 did not. Of the three paralogs, only the Group IVA PLA_2 shows enhanced activity in the presence of PIP_2, which enables its differential detection in cell homogenates. RT-PCR was used to demonstrate the presence of all three enzymes in human U937 and human WISH cells, while only Group IVA and Group IVB PLA_2 were detected in tnurine P3S8D_1 cells and human astrocytes at the mRNA level.
机译:使用杆状病毒表达系统,在Sf9昆虫细胞中过表达了三个已知的人类IV型磷脂酶A_2(PLA_2)的同系物IVA,1VB和IVC组。在昆虫细胞裂解液中鉴定出内源性,不依赖钙的PLA_2活性,可被溴烯醇内酯(BEL)抑制。 IV组PLA_2酶的特征是在BEL的存在下过表达昆虫细胞裂解物,从而使其能够与内源PLA_2活性区分开。发现IVC PLA_2组具有显着的溶血磷脂酶活性,而IVB PLA_2组则没有。在这三个旁系同源物中,只有IVA PLA_2组在PIP_2存在下显示出增强的活性,这使其能够在细胞匀浆中进行差异检测。 RT-PCR用于证明人U937和人WISH细胞中所有三种酶的存在,而在牛P3S8D_1细胞和人星形胶质细胞中仅在mRNA水平检测到IVA组和IVB PLA_2组。

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