Christian May, Regina Preisig-Müller, Michaela Höhne, Petra Gnau, Helmut Kindl
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引用次数: 59
Abstract
A patatin-like protein is present in the storage tissue of cucumber seedlings during the stage of fat mobilization. The cucumber protein is a homologue of a glycoprotein which in potatoes accounts for most of the total protein content of tubers. Following preparation of a cucumber cDNA library representing the developmental stage of cotyledons of 1 day old germinating seeds we isolated and characterized a clone encoding a patatin-like protein. Antibodies raised against the protein expressed in bacteria were used for immunodetection in subcellular fractions of cucumber seedlings. It was shown that the patatin-like protein was virtually exclusively confined to lipid bodies. The protein expressed in bacteria was characterized in vitro by its esterase activity acting on monoacylglycerols and phospholipids. Detailed analysis using various forms of phosphatidyl choline as substrates demonstrated that the patatin-like protein is a phospholipase A2 acting on palmitoyl, linoleoyl and hydroperoxidized linoleoyl groups equally well. Studying the temporal and tissue-specific expression of patatin-like protein mRNA we showed its appearance exclusively during fat catabolism. As maximal amounts of the protein were found at an early stage of fat mobilization and confined to lipid bodies, we propose that the patatin-like hydrolase is involved in lipid body mobilization.
在脂肪动员阶段,黄瓜幼苗的储存组织中存在一种patatin样蛋白。黄瓜蛋白是马铃薯中占块茎总蛋白含量大部分的糖蛋白的同源物。在制备了代表1天萌发种子子叶发育阶段的黄瓜cDNA文库之后,我们分离并鉴定了一个编码patatin样蛋白的克隆。针对细菌中表达的蛋白产生抗体,用于黄瓜幼苗亚细胞部分的免疫检测。结果表明,patatin样蛋白几乎完全局限于脂质体。体外通过对单酰基甘油和磷脂的酯酶活性对细菌表达的蛋白进行了表征。利用各种形式的磷脂酰胆碱作为底物进行详细分析,表明patatin样蛋白是一种磷脂酶A2,对棕榈酰、亚油酰和氢过氧化亚油酰都有很好的作用。通过研究patatin-like protein mRNA的时间和组织特异性表达,我们发现它只在脂肪分解代谢过程中出现。由于在脂肪动员的早期发现了最大数量的蛋白质,并且局限于脂质体,我们提出patatin样水解酶参与脂质体动员。