Hydrolysis of a fluorescent phospholipid substrate by phospholipase A2 and lipoprotein lipase.
Hydrolysis of a fluorescent phospholipid substrate by phospholipase A2 and lipoprotein lipase.
复制标题
磷脂酶 A2 和脂蛋白脂肪酶水解荧光磷脂底物。
DOI：
10.1016/0006-291x(84)91479-7
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发表时间：
1984
影响因子：
3.1
通讯作者：
Johnson,JD
中科院分区：
文献类型：
--
作者：
Wittenauer,LA;Shirai,K;Jackson,RL;Johnson,JD
以荧光磷脂1-酰基-2-[6-[（7-硝基-2，1，3-苯并恶二唑-4-基）氨基]-己酰基]磷脂酰胆碱（C6-NBD-PC）为底物，研究了猪胰磷脂酶A2（PA 2）和牛乳脂蛋白脂酶（LpL）的酶活。C6-NBD-PC的水解通过任一酶导致大于50倍的荧光增强，在540 nm处的发射最大值没有偏移; Ca++是PA 2催化所需的。对水解产物的鉴定表明，LpL和PA 2分别在sn-1和sn-2位置裂解。对于PA 2，但不是LpL，有一个显着的增强酶催化在脂质浓度高于临界胶束浓度的脂质。此外，载脂蛋白C-II，激活蛋白的LpL长链脂肪酰基底物，没有提高催化率的水溶性荧光磷脂的酶。
Abstract The fluorescent phospholipid 1-acyl-2-[6-[(7-nitro-2, 1, 3benzoxadiazol-4-yl) amino]-caproyl] phosphatidylcholine (C 6-NBD-PC) was used as a substrate for porcine pancreatic phospholipase A 2 (PA 2) and bovine milk lipoprotein lipase (LpL). Hydrolysis of C 6-NBD-PC by either enzyme resulted in a greater than 50-fold fluorescence enhancement with no shift in the emission maximum at 540 nm; Ca++ was required for PA 2 catalysis. Identification of the products of hydrolysis showed cleavage at the sn-1 and sn-2 positions for LpL and PA 2, respectively. For PA 2, but not for LpL, there was a marked enhancement of enzyme catalysis at lipid concentrations above the critical micellar concentration of the lipid. Furthermore, apolipoprotein C-II, the activator protein of LpL for long-chain fatty acyl substrates, did not enhance the rate of catalysis of the water-soluble fluorescent phospholipid for either enzyme.