Puri V, Arora A, Gupta C M
Division of Membrane Biology, Central Drug Research Institute, Lucknow, India.
Eur J Biochem. 1999 Feb;259(3):586-91. doi: 10.1046/j.1432-1327.1999.00102.x.
To understand the role of the ester moiety of the sn-1 acyl chain in phospholipase A2-glycerophospholipid interactions, we introduced an additional methylene residue between the glycerol C1 and C2 carbon atoms of phosphatidylcholines, and then studied the kinetics of hydrolysis and the binding of such butanetriol-containing phospholipids with Naja naja phospholipase A2. Hydrolysis was monitored by using phospholipids containing a NBD-labelled sn-2 acyl chain and binding was ascertained by measuring the protein tryptophan fluorescence. The hydrolysis of butanetriol-containing phospholipids was invariably slower than that of the glycerol-containing phospholipids. In addition, the enzyme binding with the substrate was markedly decreased upon replacing the glycerol residue with the 1,3,4-butanetriol moiety in phosphatidylcholines. These results have been interpreted to suggest that the sn-1 ester group in glycerophospholipids could play an important role in phospholipase A2-phospholipid interactions.
为了解磷脂酶A2与甘油磷脂相互作用中sn-1酰基链酯部分的作用，我们在磷脂酰胆碱的甘油C1和C2碳原子之间引入了一个额外的亚甲基残基，然后研究了含丁三醇的磷脂的水解动力学以及此类磷脂与眼镜蛇磷脂酶A2的结合。通过使用含有NBD标记的sn-2酰基链的磷脂监测水解，并通过测量蛋白质色氨酸荧光确定结合情况。含丁三醇的磷脂的水解总是比含甘油的磷脂慢。此外，在磷脂酰胆碱中用1,3,4-丁三醇部分取代甘油残基后，酶与底物的结合明显减少。这些结果被解释为表明甘油磷脂中的sn-1酯基在磷脂酶A2-磷脂相互作用中可能起重要作用。
