The x-ray structure of Brassica napus β-keto acyl carrier protein reductase and its implications for substrate binding and catalysis

Fisher, M. and Kroon, J.T.M. and Martindale, Wayne and Stuitje, A.R. and Slabas, A.R. and Rafferty, J.B. (2000) The x-ray structure of Brassica napus β-keto acyl carrier protein reductase and its implications for substrate binding and catalysis. Structure, 8 (4). pp. 339-347. ISSN 09692126

Full content URL: https://www.scopus.com/inward/record.uri?eid=2-s2....

Documents
The x-ray structure of Brassica napus β-keto acyl carrier protein reductase and its implications for substrate binding and catalysis

Request a copy
[img] PDF
1-s2.0-S0969212600001155-main.pdf__tid=9c3b9b4c-a7e5-49c0-a2ad-2bc92495470b&acdnat=1542893296_212834d4557f9e865ac899c4fc7d1052 - Whole Document
Restricted to Repository staff only
Available under License Creative Commons Attribution-NonCommercial 4.0 International.

556kB
Item Type:Article
Item Status:Live Archive

Abstract

Background: β-Keto acyl carrier protein reductase (BKR) catalyzes the pyridine-nucleotide-dependent reduction of a 3-oxoacyl form of acyl carrier protein (ACP), the first reductive step in de novo fatty acid biosynthesis and a reaction often performed in polyketide biosynthesis. The Brassica napus BKR enzyme is NADPH-dependent and forms part of a dissociable type II fatty acid synthetase (FAS). Significant sequence similarity is observed with enoyl acyl carrier protein reductase (ENR), the other reductase of FAS, and the short-chain alcohol dehydrogenase (SDR) family. Results: The first crystal structure of BKR has been determined at 2.3 à resolution in a binary complex with an NADP+ cofactor. The structure reveals a homotetramer in which each subunit has a classical dinucleotide-binding fold. A triad of Ser154, Tyr167 and Lys171 residues is found at the active site, characteristic of the SDR family. Overall BKR has a very similar structure to ENR with good superimposition of catalytically important groups. Modelling of the substrate into the active site of BKR indicates the need for conformational changes in the enzyme. Conclusions: A catalytic mechanism can be proposed involving the conserved triad. Helix α6 must shift its position to permit substrate binding to BKR and might act as a flexible lid on the active site. The similarities in fold, mechanism and substrate binding between BKR, which catalyzes a carbon-oxygen double-bond reduction, and ENR, the carbon-carbon double-bond oxidoreductase in FAS, suggest a close evolutionary link during the development of the fatty acid biosynthetic pathway.

Additional Information:cited By 81
Keywords:acyl carrier protein, alcohol dehydrogenase, fatty acid synthase, ferredoxin nicotinamide adenine dinucleotide phosphate reductase, oxidoreductase, article, binding site, biosynthesis, Brassica, catalysis, enzyme activity, enzyme structure, enzyme substrate, Escherichia coli, nonhuman, priority journal, X ray diffraction, Brassica, Brassica napus
Subjects:C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:College of Science > National Centre for Food Manufacturing
ID Code:33772
Deposited On:27 Nov 2018 12:25

Repository Staff Only: item control page