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球形马拉色菌脂肪酶基因LIP1的分离与表达

Isolation and expression of a Malassezia globosa lipase gene, LIP1.

作者信息

DeAngelis Yvonne M, Saunders Charles W, Johnstone Kevin R, Reeder Nancy L, Coleman Christal G, Kaczvinsky Joseph R, Gale Celeste, Walter Richard, Mekel Marlene, Lacey Martin P, Keough Thomas W, Fieno Angela, Grant Raymond A, Begley Bill, Sun Yiping, Fuentes Gary, Youngquist R Scott, Xu Jun, Dawson Thomas L

机构信息

1The Procter & Gamble Company, Miami Valley Innovation Center, Cincinnati, Ohio, USA.

出版信息

J Invest Dermatol. 2007 Sep;127(9):2138-46. doi: 10.1038/sj.jid.5700844. Epub 2007 Apr 26.

DOI:10.1038/sj.jid.5700844
PMID:17460728
Abstract

Dandruff and seborrheic dermatitis (D/SD) are common hyperproliferative scalp disorders with a similar etiology. Both result, in part, from metabolic activity of Malassezia globosa and Malassezia restricta, commensal basidiomycete yeasts commonly found on human scalps. Current hypotheses about the mechanism of D/SD include Malassezia-induced fatty acid metabolism, particularly lipase-mediated breakdown of sebaceous lipids and release of irritating free fatty acids. We report that lipase activity was detected in four species of Malassezia, including M. globosa. We isolated lipase activity by washing M. globosa cells. The isolated lipase was active against diolein, but not triolein. In contrast, intact cells showed lipase activity against both substrates, suggesting the presence of at least another lipase. The diglyceride-hydrolyzing lipase was purified from the extract, and much of its sequence was determined by peptide sequencing. The corresponding lipase gene (LIP1) was cloned and sequenced. Confirmation that LIP1 encoded a functional lipase was obtained using a covalent lipase inhibitor. LIP1 was differentially expressed in vitro. Expression was detected on three out of five human scalps, as indicated by reverse transcription-PCR. This is the first step in a molecular description of lipid metabolism on the scalp, ultimately leading toward a test of its role in D/SD etiology.

摘要

头皮屑和脂溢性皮炎(D/SD)是常见的过度增殖性头皮疾病,病因相似。二者部分是由球形马拉色菌和限制马拉色菌的代谢活动引起的,这两种共生担子菌酵母常见于人类头皮上。目前关于D/SD发病机制的假说包括马拉色菌诱导的脂肪酸代谢,特别是脂肪酶介导的皮脂脂质分解和刺激性游离脂肪酸的释放。我们报告在包括球形马拉色菌在内的四种马拉色菌中检测到了脂肪酶活性。我们通过洗涤球形马拉色菌细胞分离出了脂肪酶活性。分离出的脂肪酶对甘油二油酸酯有活性,但对三油酸甘油酯无活性。相比之下,完整细胞对两种底物都显示出脂肪酶活性,这表明至少存在另一种脂肪酶。从提取物中纯化出了水解甘油二酯的脂肪酶,并通过肽测序确定了其大部分序列。克隆并测序了相应的脂肪酶基因(LIP1)。使用共价脂肪酶抑制剂证实LIP1编码一种功能性脂肪酶。LIP1在体外有差异表达。逆转录-聚合酶链反应表明,在五个人类头皮样本中有三个检测到了其表达。这是对头皮脂质代谢进行分子描述的第一步,最终将对其在D/SD病因学中的作用进行检验。

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