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兔胰腺甘油三酯脂肪酶的分子克隆与特性分析

Molecular cloning and characterization of rabbit pancreatic triglyceride lipase.

作者信息

Aleman-Gomez J A, Colwell N S, Sasser T, Kumar V B

机构信息

Geriatric Research Education and Clinical Center, VA Medical Center, St. Louis, MO 63125.

出版信息

Biochem Biophys Res Commun. 1992 Nov 16;188(3):964-71. doi: 10.1016/0006-291x(92)91326-l.

Abstract

Pancreatic lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) has been cloned from a gt11 cDNA library made from poly A+ RNA of adult rabbit pancreas. Pancreatic lipase (PL) assists the absorption of dietary triglycerides by hydrolyzing them at 1 and 3 positions to free fatty acids and 2-monoacylglycerol in the presence of bile acids and colipase in the intestinal lumen. Since rabbits are classifically used for the study of the diet induced changes in the lipid metabolism, as a prelude to studying the diet and age dependent changes in the expression of this enzyme, a full length PL cDNA clone was obtained from its pancreas. The coding region of rabbit pancreatic lipase cDNA consists of 1407 base pairs contained in an open reading frame encoding 469 amino acids including the 16 that constitute the signal peptide. Northern blot analysis revealed a band around 1.5 kb. When rabbit enzyme is compared to other species, an over all homology of 70-80% was observed at the nucleotide level. High homology in the amino acid sequence and composition is also apparent between rabbit and other species like dog (65%), pig (76%) and rat (63%). Highest homology is found to be around active-site serine. The regions of homology with other species may help to define sites of interaction of lipase with co-lipase.

摘要

胰腺脂肪酶(三酰甘油酰基水解酶,EC 3.1.1.3)已从一个由成年兔胰腺的聚腺苷酸加RNA构建的gt11 cDNA文库中克隆出来。胰腺脂肪酶(PL)在肠腔内胆汁酸和辅脂酶存在的情况下,通过将膳食甘油三酯在1位和3位水解为游离脂肪酸和2-单酰甘油,来协助膳食甘油三酯的吸收。由于兔子常用于饮食诱导的脂质代谢变化的研究,作为研究该酶表达的饮食和年龄依赖性变化的前奏,从其胰腺中获得了一个全长PL cDNA克隆。兔胰腺脂肪酶cDNA的编码区由1407个碱基对组成,包含在一个开放阅读框中,该开放阅读框编码469个氨基酸,包括构成信号肽的16个氨基酸。Northern印迹分析显示一条约1.5 kb的条带。当将兔酶与其他物种进行比较时,在核苷酸水平上观察到总体同源性为70 - 80%。兔与其他物种如狗(65%)、猪(76%)和大鼠(63%)之间在氨基酸序列和组成上也具有高度同源性。发现最高同源性位于活性位点丝氨酸周围。与其他物种的同源区域可能有助于确定脂肪酶与辅脂酶相互作用的位点。

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