Biochemical characterization of the vacuolar palmitoyl acyltransferase

被引：10

作者：

Veit, M

Dietrich, LEP

Ungermann, C

机构：

[1] Univ Heidelberg, Zentrum Biochem, D-69120 Heidelberg, Germany

[2] Free Univ Berlin, Dept Immunol & Mol Biol, Fac Vet Med, D-10115 Berlin, Germany

来源：

FEBS LETTERS | 2003年 / 540卷 / 1-3期

关键词：

vacuole fusion; Vac8; palmitoylation; SNARE complex; Sec18;

D O I：

10.1016/S0014-5793(03)00232-1

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Vacuole fusion requires Sec18p-dependent acylation of the armadillo-repeat protein Vac8p that has been isolated with cis-SNARE complexes. To gain more insight into the mechanism of acylation, we analyzed the palmitoylation reaction on isolated vacuoles or in vacuolar detergent extracts. Recombinant Vac8p is palmitoylated when added to vacuoles and is anchored to membranes after modification. The palmitoyl acyltransferase (PAT) extracted from vacuolar membranes is functional in detergent extracts and shows all characteristics of an enzymatic activity: It modifies exogenous Vac8p in a temperature-, dose- and time-dependent manner, and is sensitive to bromo-palmitate, a known inhibitor of protein palmitoylation in vivo. Importantly, PAT is specific for palmitoyl-CoA, since myristoyl- and stearyl-CoA can compete with labeled Pal-CoA only at 10-fold higher amounts. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

引用

页码：101 / 105

页数：5

共 40 条

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