title: Structure of conjugated polyketone reductase from Candida parapsilosis IFO 0708 reveals conformational changes for substrate recognition upon NADPH binding.
abstract: Conjugated polyketone reductase C2 (CPR-C2) from Candida parapsilosis IFO 0708, identified as a nicotinamide adenine dinucleotide phosphate (NADPH)-dependent ketopantoyl lactone reductase, belongs to the aldo-keto reductase superfamily. This enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner. To elucidate the structural basis of the substrate specificity, we determined the crystal structures of the apo CPR-C2 and CPR-C2/NADPH complex at 1.70 and 1.80 Å resolutions, respectively. CPR-C2 adopted a triose-phosphate isomerase barrel fold at the core of the structure. Binding with the cofactor NADPH induced conformational changes in which Thr27 and Lys28 moved 15 and 5.0 Å, respectively, in the close vicinity of the adenosine 2'-phosphate group of NADPH to form hydrogen bonds. Based on the comparison of the CPR-C2/NADPH structure with 3-α-hydroxysteroid dehydrogenase and mutation analyses, we constructed substrate binding models with ketopantoyl lactone, which provided insight into the substrate specificity by the cofactor-induced structure. The results will be useful for the rational design of CPR-C2 mutants targeted for use in the industrial manufacture of ketopantoyl lactone.
authors.affiliation: Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: Qin
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yamamura
authors.forename: Takuya
authors.initials: T
authors.lastname: Miyakawa
authors.forename: Michihiko
authors.initials: M
authors.lastname: Kataoka
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nagai
authors.forename: Nahoko
authors.initials: N
authors.lastname: Kitamura
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Urano
authors.forename: Shintaro
authors.initials: S
authors.lastname: Maruoka
authors.forename: Jun
authors.initials: J
authors.lastname: Ohtsuka
authors.forename: Koji
authors.initials: K
authors.lastname: Nagata
authors.forename: Sakayu
authors.initials: S
authors.lastname: Shimizu
authors.forename: Masaru
authors.initials: M
authors.lastname: Tanokura
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000378:metabolism*; D002175:Candida / Q000201:enzymology*; D018360:Crystallography, X-Ray; D004252:DNA Mutational Analysis; D008958:Models, Molecular; D009249:NADP / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: C008232:keto-pantoyllactone; D009249:NADP; C005412:pantolactone; D000429:Alcohol Oxidoreductases; C021935:2-oxopantoyl lactone reductase; D015107:4-Butyrolactone