BIOSYNTHESIS OF PHOSPHOINOSITOL-CONTAINING SPHINGOLIPIDS FROM PHOSPHATIDYLINOSITOL BY A MEMBRANE PREPARATION FROM SACCHAROMYCES-CEREVISIAE
BIOSYNTHESIS OF PHOSPHOINOSITOL-CONTAINING SPHINGOLIPIDS FROM PHOSPHATIDYLINOSITOL BY A MEMBRANE PREPARATION FROM SACCHAROMYCES-CEREVISIAE
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DOI：
10.1128/jb.142.3.747-754.1980
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发表时间：
1980-01-01
影响因子：
3.2
通讯作者：
LESTER, RL
中科院分区：
文献类型：
--
作者：
BECKER, GW;LESTER, RL
从S.用[32 P]磷脂酰[3 H]肌醇处理酿酒酵母，导致两种标记转移到2种产物，其被表征为2种属。肌醇磷酸神经酰胺，在分子的神经酰胺部分不同。产物的特征在于在弱碱中的稳定性，在硅胶浸渍纸上的流动性，在硫酸柱上的色谱和在碱水解时释放肌醇磷酸的基础上。该反应不需要添加金属，也不受EDTA抑制。洗涤剂Triton X-100和吐温20提供很少的刺激，如果有的话。在相对高浓度的磷脂酰肌醇（1-4 mM）下，体外速率约为体内速率的20%。虽然神经酰胺是一个合乎逻辑的底物，该反应不能大大刺激通过添加含有单和二羟基脂肪酸的神经酰胺。此外，酵母膜与[32 P]磷脂酰肌醇孵育产生了一种产物，该产物在色谱上与主要的酵母磷脂，甘露糖-（肌醇-P）2神经酰胺无法区分。
Incubation of membranes prepared from S. cerevisiae with [32P]phosphatidyl[3H]inositol resulted in the transfer of both labels to 2 products which were characterized as 2 spp. of inositolphosphoceramide, differing in the ceramide portion of the molecule. The products were characterized on the basis of stability in mild alkali, mobility on silica gel-impregnated paper, chromatography on silicic acid columns and release of inositol phosphate upon base hydrolysis. The reaction did not require the addition of metals, nor was it inhibited by EDTA. The detergents Triton X-100 and Tween 20 provided little, if any, stimulation. At relatively high concentrations of phosphatidylinositol (1-4 mM), the in vitro rate was about 20% of the in vivo rate. Although ceramide was a logical substrate, the reaction could not be greatly stimulated by the addition of ceramides containing mono- and dihydroxy fatty acids. In addition, incubation of yeast membranes with [32P]phosphatidylinositol gave rise to a product that was chromatographically indistinguishable from the major yeast phosphosphingolipid, mannose-(inositol-P)2 ceramide.