Kinetic mechanism of NADH-enoyl-ACP reductase from Brassica napus.
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.
Fawcett
T
T
Department of Biological Sciences, University of Durham, DH1 3LE, Durham, UK. tony.fawcett@durham.ac.uk
Copse
C L
CL
Simon
J W
JW
Slabas
A R
AR
D001667:Binding, Competitive; D001937:Brassica / Q000201:enzymology*; D002384:Catalysis; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH); D004795:Enzyme Stability; D007700:Kinetics; D008970:Molecular Weight; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
D011994:Recombinant Proteins; D010088:Oxidoreductases; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH)