Characterization of pancreatic lipase-related protein 2 isolated from human pancreatic juice
Characterization of pancreatic lipase-related protein 2 isolated from human pancreatic juice
复制标题
DOI：
10.1016/j.bbapap.2004.06.005
复制
发表时间：
2004-09-01
影响因子：
3.2
通讯作者：
De Caro, A
中科院分区：
文献类型：
--
作者：
De Caro, J;Sias, B;De Caro, A
人胰脂肪酶相关蛋白2（HPLRP 2）首次在胰液中使用特异性抗肽抗体鉴定并纯化至均一。使用来自cDNA推导的HPLRP 2蛋白序列的合成肽在兔中产生抗体。Western印迹分析表明，这些抗体不与经典的人胰脂肪酶（HPL）或人胰脂肪酶相关蛋白I（HPLRP 1），但与天然大鼠PLRP 2（RPLRP 2），以及与重组大鼠和豚鼠PLRP 2（GPLRP 2）交叉反应。对固定化的抗重组HPLRP 2多克隆抗体进行免疫亲和层析，以在常规层析步骤（包括凝胶过滤和阴离子交换剂上的层析）后纯化天然HPLRP 2。使用各种甘油三酯、磷脂和半乳糖脂作为底物研究HPLRP 2的底物特异性。胆盐可抑制脂肪酶对甘油三酯的催化活性，辅脂酶可使其微弱地恢复。HPLRP 2在磷脂胶束上的磷脂酶活性很低。一个显着水平的半乳糖脂酶活性测定使用单半乳糖甘油二酯单分子膜。这些数据表明，HPLRP 2的主要生理功能是水解半乳糖脂，这是植物性食品中存在的主要脂质。(C)2004 Elsevier B.V.保留所有权利。
Human pancreatic lipase-related protein 2 (HPLRP2) was identified for the first time in pancreatic juice using specific anti-peptide antibodies and purified to homogeneity. Antibodies were raised in the rabbit using a synthetic peptide from the HPLRP2 protein sequence deduced from cDNA. Western blotting analysis showed that these antibodies did not react with classical human pancreatic lipase (HPL) or human pancreatic lipase-related protein I (HPLRP1) but cross-reacted with native rat PLRP2 (RPLRP2), as well as with recombinant rat and guinea-pig PLRP2 (GPLRP2). Immunoaffinity chromatography was performed on immobilized anti-recombinant HPLRP2 polyclonal antibodies to purify native HPLRP2 after conventional chromatographic steps including gel filtration and chromatrography on an anion-exchanger. The substrate specificity of HPLRP2 was investigated using various triglycerides, phospholipids and galactolipids as substrates. The lipase activity on triglycerides was inhibited by bile salts and weakly restored by colipase. The phospholipase activity of HPLRP2 on phospholipid micelles was very low. A significant level of galactolipase activity was measured using monogalactosyldiglyceride monomolecular films. These data suggest that the main physiological function of HPLRP2 is the hydrolysis of galactolipids, which are the main lipids present in vegetable food. (C) 2004 Elsevier B.V. All rights reserved.