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Kinetic mechanism of NADH‐enoyl‐ACP reductase from Brassica napus
Author(s) -
Fawcett Tony,
Copse Catherine L,
Simon J.William,
Slabas Antoni R
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)02128-1
Subject(s) - reductase , chemistry , substrate (aquarium) , enzyme , ternary complex , biochemistry , mass spectrometry , atp synthase , stereochemistry , chromatography , biology , ecology
Enoyl‐ACP reductase, a component of fatty acid synthase, is a target for anti‐microbial agents and herbicides. Here we demonstrate the kinetic mechanism to be a compulsory‐order ternary complex with NADH binding before the acyl substrate. Matrix‐assisted laser desorption ionisation mass spectrometry analysis of enzymatically and synthesised crotonyl‐ACP substrate showed the former to contain a single acyl group, whereas the latter contained up to four additional crotonylations. The use of authentic crotonyl‐ACP will be important in future kinetic and crystallographic studies.