title: Crystallization and preliminary X-ray crystallographic studies of enoyl-acyl carrier protein reductase (FabI) from Psuedomonas aeruginosa.
abstract: During fatty-acid biosynthesis, enoyl-acyl carrier protein (enoyl-ACP) reductase catalyzes the reduction of trans-2-enoyl-ACP to fully saturated acyl-ACP via the ubiquitous fatty-acid synthase system. NADH-dependent enoyl-ACP reductase (FabI) from Pseudomonas aeruginosa has been purified and crystallized as an apoenzyme and in a complex form with NADH and triclosan. Triclosan is an inhibitor of FabI and forms a stable ternary complex in the presence of NADH. The crystals of native and complexed FabI diffracted to resolutions of 2.6 and 1.8 Å, respectively. The crystals both belonged to space group P2(1), with unit-cell parameters a = 117.32, b = 155.844, c = 129.448 Å, β = 111.061° for the native enzyme and a = 64.784, b = 107.573, c = 73.517 Å, β = 116.162° for the complex. Preliminary molecular replacement further confirmed the presence of four tetramers of native FabI and one tetramer of the complex in the asymmetric unit, corresponding to Matthews coefficients (V(M)) of 2.46 and 2.05 Å(3) Da(-1) and solvent contents of 50.1 and 40.1%, respectively.
authors.affiliation: Division of Biotechnology, College of Life Sciences, Korea University, Seoul, Republic of Korea.
authors.forename: Jeong Hye
authors.initials: JH
authors.lastname: Lee
authors.forename: Ae Kyung
authors.initials: AK
authors.lastname: Park
authors.forename: Young Min
authors.initials: YM
authors.lastname: Chi
authors.forename: Jin Ho
authors.initials: JH
authors.lastname: Moon
authors.forename: Ki Seog
authors.initials: KS
authors.lastname: Lee
mesh_terms: D001051:Apoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH) / Q000737:chemistry*; D064429:Fatty Acid Synthases / Q000378:metabolism; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000378:metabolism; D014260:Triclosan / Q000378:metabolism / Q000494:pharmacology; D014961:X-Ray Diffraction
chemical_list: D001051:Apoenzymes; D001426:Bacterial Proteins; D009243:NAD; D014260:Triclosan; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH); D064429:Fatty Acid Synthases