Identification of major proteins in the lipid droplet-enriched fraction isolated from the human hepatocyte cell line HuH7
Identification of major proteins in the lipid droplet-enriched fraction isolated from the human hepatocyte cell line HuH7
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DOI：
10.1016/j.bbamcr.2003.10.018
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发表时间：
2004-02-02
影响因子：
5.1
通讯作者：
Takano, T
中科院分区：
文献类型：
--
作者：
Fujimoto, Y;Itabe, H;Takano, T
最近的研究已经揭示了细胞内脂滴在多种物种中的存在。在哺乳动物细胞中，存在特异性定位于脂滴中的蛋白质。然而，液滴中的蛋白质谱仍有待澄清。在这项研究中，富含脂滴的馏分分离从人肝细胞系HuH 7使用蔗糖密度梯度离心，和17个主要蛋白质的馏分中使用纳米LC-MS/MS技术进行了鉴定。脂肪分化相关蛋白（ADRP）是组分中含量最高的蛋白质。第二丰富蛋白被鉴定为酰基辅酶A合成酶3（ACS 3）和17 β-羟基类固醇脱氢酶11（17 β HSD 11）。包括在确定的蛋白质是五个脂质代谢酶以及两个脂滴特异性蛋白质。当HuH 7细胞裂解物通过密度梯度分级时，在液滴富集级分中发现大部分17 β HSD 11。在免疫细胞化学分析中，17 β HSD 11显示与ADRP重叠的环形图像。这些结果表明，一组特定的蛋白质富集在富含脂滴的馏分中，并且17 β HSD 11特异性地定位在该馏分中。(C)2003 Elsevier B. V.保留所有权利。
Recent studies have revealed the presence of intracellular lipid droplets in wide variety of species. In mammalian cells, there exist proteins specifically localize in lipid droplets. However, the protein profile in the droplet remains yet to be clarified. In this study, a fraction enriched with lipid droplets was isolated from a human hepatocyte cell line HuH7 using sucrose density gradient centrifugation, and 17 major proteins in the fraction were identified using nano LC-MS/MS techniques. Adipose differentiation-related protein (ADRP) was the most abundant protein in the fraction. The secondary abundant proteins were identified to be acyl-CoA synthetase 3 (ACS3) and 17beta-hydroxysteroid dehydrogenase 11 (17 betaHSD11). Included in the identified proteins were five lipid-metabolizing enzymes as well as two lipid droplet-specific proteins. When HuH7 cell lysate was fractionated by a density gradient, most of 17betaHSD11 was found in the droplet-enriched fraction. In immunocytochemical analysis, 17betaHSD11 showed ring-shaped images which overlapped with those for ADRP. These results suggest that a specific set of proteins is enriched in the lipid droplet-enriched fraction and that 17betaHSD11 localizes specifically in the fraction. (C) 2003 Elsevier B.V. All rights reserved.