RESPONSES OF PURIFIED PHOSPHOLIPASE-A(2) TO PHOSPHOLIPASE-A(2) ACTIVATING PROTEIN (PLAP) AND MELITTIN
RESPONSES OF PURIFIED PHOSPHOLIPASE-A(2) TO PHOSPHOLIPASE-A(2) ACTIVATING PROTEIN (PLAP) AND MELITTIN
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DOI：
10.1016/0005-2760(93)90292-h
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发表时间：
1993-02-10
期刊：
影响因子：
--
通讯作者：
CLARK, MA
中科院分区：
文献类型：
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作者：
STEINER, MR;BOMALASKI, JS;CLARK, MA
通过测定 PLAP 对两种纯化 PLA2 的影响来评估磷脂酶 A2 (PLA2) 刺激蛋白 PLAP 在 PLA2 活性调节中的作用。大约。通过阳离子交换 HPLC 和免疫亲和 HPLC（使用针对炎症渗出物 PLA2 的 N 末端序列的抗血清）从小鼠胸腺瘤细胞 EL-4 细胞中纯化 14 kDa 酶。大约。通过连续疏水、阴离子交换、羟基磷灰石和凝胶过滤 HPLC，从小鼠乳腺癌衍生细胞中纯化出 110 kDa 酶。 PLAP 和蜂毒肽（一种来自蜂毒的免疫相关 PLA2 刺激肽）都不会增加这种高分子量酶的活性。相比之下，PLAP 对低分子量 PLA2 的刺激超过 20 倍，大约 10 倍。蜂毒肽的 5 倍刺激。当蛋白质与磷脂的比例为1:10(6)时，观察到PLAP对酶活性的刺激，而蜂毒肽与磷脂的比例为1:3。因此，PLAP介导的PLA2活性刺激可能包括PLAP和酶之间的相互作用，与蜂毒肽刺激相反，蜂毒肽刺激涉及蜂毒肽和磷脂之间的相互作用。
The role of the phospholipase A2 (PLA2) stimulating protein PLAP in the regulation of PLA2 activity was assessed by determination of the effects of PLAP on two purified PLA2s. An approx. 14 kDa enzyme was purified from mouse thymoma cells, EL-4 cells, by cation ion exchange HPLC and immunoaffinity HPLC (with antiserum to the N-terminal sequence of an inflammatory exudate PLA2). An approx. 110 kDa enzyme was purified from mouse mammary carcinoma derived cells by sequential hydrophobic, anion exchange, hydroxyapatite and gel filtration HPLC. Neither PLAP nor melittin, an immunologically related PLA2 stimulating peptide from bee venom, increased the activity of the high molecular weight enzyme. In contrast, there was more than a 20-fold stimulation of the low molecular weight PLA2 by PLAP and an approx. 5-fold stimulation by melittin. The stimulation of enzyme activity by PLAP was observed at a protein to phospholipid ratio of 1:10(6) while the ratio of melittin to phospholipid was 1:3. Thus, PLAP mediated stimulation of PLA2 activity may include an interaction between PLAP and the enzyme, in contrast to melittin stimulation, which involves interactions between melittin and phospholipid.