四膜虫的蛋白质前酰化

Phillip E. Ryals
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引用次数: 2

摘要

检测了vorax四膜虫V2S和梨状四膜虫GL中是否存在甲戊酸衍生脂质共价修饰的蛋白质。用[3H]甲羟戊酸内酯、[3H]甲羟戊酸酯、[3H]法尼基焦磷酸或[3H]香叶香叶基焦磷酸对细胞进行放射性标记,在丙酮中蛋白质脱脱后出现与蛋白质相关的放射性。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳标记的蛋白质显示放射性多肽的分子量范围从123到14 kDa的凝胶切片闪烁计数或荧光。对总标记蛋白进行碘化甲基裂解,随后进行HPLC分析和释放的戊烯醇的闪烁计数,显示与市售的法尼醇(C15)和香叶醇(C20)共层析的放射性峰。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Protein Prenylation in Tetrahymena

Tetrahymena vorax V2S and Tetrahymena pyriformis GL were examined for the presence of proteins covalently modified with mevalonate-derived lipids. Radiolabeling of cells using [3H]mevalonolactone, [3H]mevalonate, [3H]farnesyl pyrophosphate or [3H]geranylgeranyl pyrophosphate resulted in the appearance of protein-associated radioactivity following protein delipidation in acetone. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of labeled protein revealed radioactive polypeptides ranging in molecular mass from 123 to 14 kDa following scintillation counting of gel slices or fluorography. Methyl iodide cleavage of total labeled protein followed by HPLC analysis and scintillation counting of the released prenyl alcohols showed radioactive peaks that co-chromatographed with commercially obtained farnesol (C15) and geranylgeraniol (C20).

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