Purification and kinetic properties of lysophosphatidylinositol acyltransferase from bovine heart muscle microsomes and comparison with lysophosphatidylcholine acyltransferase.
Purification and kinetic properties of lysophosphatidylinositol acyltransferase from bovine heart muscle microsomes and comparison with lysophosphatidylcholine acyltransferase.
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牛心肌微粒体溶血磷脂酰肌醇酰基转移酶的纯化和动力学特性以及与溶血磷脂酰胆碱酰基转移酶的比较。
DOI：
10.1016/0003-9861(89)90290-7
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发表时间：
1989
影响因子：
3.9
通讯作者：
R. MacQuarrie
中科院分区：
文献类型：
--
作者：
M. Sanjanwala;Grace Y. Sun;R. MacQuarrie
酰基-CoA:1-酰基-sn-甘油-3-磷酸肌醇酰基转移酶（LPI 酰基转移酶，EC 2.3.1.23）从牛心肌微粒体纯化约 11,000 倍至接近同质。通过用洗涤剂 3-((3-胆酰胺基丙基)二甲基铵)-1-丙磺酸盐提取，然后在 Cibacron 蓝色琼脂糖、DEAE-纤维素和 Matrex 凝胶绿 A 上进行色谱法进行纯化。在十二烷基硫酸盐存在下通过聚丙烯酰胺凝胶电泳测量，分离的酶是 58,000 Da 的单一蛋白质。该纯化程序还可以分离相关的溶血磷脂酰胆碱 (LPC) 酰基转移酶，该酶在最后的色谱步骤中与 LPI 酰基转移酶分离。纯化的 LPI 酰基转移酶对 LPI 作为酰基受体表现出绝对特异性。尽管优选的底物是长链不饱和衍生物，但发现酰基辅酶 A 衍生物作为底物具有更广泛的特异性：测量的反应性顺序为花生四烯酰辅酶 A > 油酰辅酶 A > 二十碳二烯酰辅酶 A > 亚油酰辅酶 A。使用棕榈酰辅酶 A 或硬脂酰辅酶 A 作为潜在底物时，几乎没有发现活性。这些特性与酶在控制磷酸肌醇的酰基组成中的作用一致。 LPC酰基转移酶和LPI酰基转移酶的比较表明，这两种酶具有不同的动力学和物理特性，并且受局麻药（作为有效抑制剂）的影响不同。
The enzyme acyl-CoA:1-acyl-sn-glycero-3-phosphoinositol acyltransferase (LPI acyltransferase, EC 2.3.1.23) was purified approximately 11,000-fold to near homogeneity from bovine heart muscle microsomes. The purification was effected by extraction with the detergent 3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate, followed by chromatography on Cibacron blue agarose, DEAE-cellulose, and Matrex gel green A. The isolated enzyme was a single protein of 58,000 Da as measured by polyacrylamide gel electrophoresis in the presence of dodecyl sulfate. This purification procedure also allows isolation of the related enzyme lysophosphatidylcholine (LPC) acyltransferase, which was separated from LPI acyltransferase at the final chromatographic step. The purified LPI acyltransferase exhibits an absolute specificity for LPI as the acyl acceptor. Broader specificity was found for acyl-CoA derivatives as substrates, although the preferred substrates are long-chain, unsaturated derivatives: measured reactivities were in the order arachidonoyl-CoA > oleoyl-CoA > eicosadienoyl-CoA > linoleoyl-CoA. Little activity was found with palmitoyl-CoA or stearoyl-CoA as potential substrates. These properties are consistent with a role of the enzyme in controlling the acyl group composition of phosphoinositides. Comparison of LPC acyltransferase and LPI acyltransferase shows that these two enzymes have distinct kinetic and physical properties and are affected differently by local anesthetics, which are potent inhibitors.
通过稳定同位素技术评估磷脂脱酰-再酰化循环。
DOI：
10.1016/0006-291x(87)90454-2
发表时间：
1987
影响因子：
3.1
作者：
Kuwae,T;Schmid,PC;Schmid,HH
通讯作者：
Schmid,HH
乙酰胆碱刺激 C62B 神经胶质瘤细胞中花生四烯酸的选择性释放和再酯化以及磷酸肌醇和甘油磷酸肌醇的积累。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
DeGeorge,JJ;Ousley,AH;McCarthy,KD;Lapetina,EG;Morell,P
通讯作者：
Morell,P