Fawcett T, Copse C L, Simon J W, Slabas A R
Department of Biological Sciences, University of Durham, DH1 3LE, Durham, UK.
FEBS Lett. 2000 Nov 3;484(2):65-8. doi: 10.1016/s0014-5793(00)02128-1.
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.
