Functional characterization of GcpE, an essential enzyme of the non-mevalonate pathway of isoprenoid biosynthesis

被引：91

作者：

Kollas, AK

Duin, EC

Eberl, M

Altincicek, B

Hintz, M

Reichenberg, A

Henschker, D

Henne, A

Steinbrecher, I

Ostrovsky, DN

Hedderich, R

Beck, E

Jomaa, H

Wiesner, J

机构：

[1] Univ Giessen, Inst Biochem, D-35392 Giessen, Germany

[2] Max Planck Inst Terr Mikrobiol, D-35043 Marburg, Germany

[3] Jomaa Pharmaka GMBH, D-35392 Giessen, Germany

[4] Univ Gottingen, Inst Mikrobiol & Genet, D-37077 Gottingen, Germany

[5] AN Bakh Biochem Inst, Moscow 119071, Russia

来源：

FEBS LETTERS | 2002年 / 532卷 / 03期

基金：

俄罗斯基础研究基金会;

关键词：

2-C-methyl-D-erythritol-2,4-cyclodiphosphate; GcpE; (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate; isoprenoid biosynthesis; LytB; 2-C-methyl-D-erythritol-4-phosphate pathway;

D O I：

10.1016/S0014-5793(02)03725-0

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：

The gcpE gene product controls one of the terminal steps of isoprenoid biosynthesis via the mevalonate independent 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway. This pathway is utilized by a variety of eubacteria, the plastids of algae and higher plants, and the plastid-like organelle of malaria parasites. Recombinant GcpE protein from the hyperthermophilic bacterium Thermus thermophilus was produced in Escherichia coli and purified under dioxygen-free conditions. The protein was enzymatically active in converting 2-C-methyl-D-erythritol-2,4-cyclodiphosphate (MEcPP) into (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP) in the presence of dithionite as reductant. The maximal specific activity was 0.6 mumol min(-1) mg(-1) at pH 7.5 and 55degreesC. The k(cat) value was 0.4 S-1 and the K-m value for HMBPP 0.42 mM. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

引用

页码：432 / 436

页数：5

共 20 条

[1] Cutting edge:: Human γδ T cells are activated by intermediates of the 2-C-methyl-D-erythritol 4-phosphate pathway of isoprenoid biosynthesis [J].