title: Crystallization of an atypical short-chain dehydrogenase from Vibrio vulnificus lacking the conserved catalytic tetrad.
abstract: Short-chain dehydrogenases/reductases (SDRs) are a rapidly expanding superfamily of enzymes that are found in all kingdoms of life. Hallmarked by a highly conserved Asn-Ser-Tyr-Lys catalytic tetrad, SDRs have a broad substrate spectrum and play diverse roles in key metabolic processes. Locus tag VVA1599 in Vibrio vulnificus encodes a short-chain dehydrogenase (hereafter referred to as SDRvv) which lacks the signature catalytic tetrad of SDR members. Structure-based protein sequence alignments have suggested that SDRvv may harbour a unique binding site for its nicotinamide cofactor. To date, structural studies of SDRs with altered catalytic centres are underrepresented in the scientific literature, thus limiting understanding of their spectrum of substrate and cofactor preferences. Here, the expression, purification and crystallization of recombinant SDRvv are presented. Two well diffracting crystal forms could be obtained by cocrystallization in the presence of the reduced form of the phosphorylated nicotinamide cofactor NADPH. The collected data were of sufficient quality for successful structure determination by molecular replacement and subsequent refinement. This work sets the stage for deriving the identity of the natural substrate of SDRvv and the structure-function landscape of typical and atypical SDRs.
authors: Unit for Structural Biology, Laboratory for Protein Biochemistry and Biomolecular Engineering, Unit for Structural Biology, Ghent University, K. L. Ledeganckstraat 35, B-9000 Ghent, Belgium.
authors: Geraldine
authors: G
authors: Buysschaert
authors: Kenneth
authors: K
authors: Verstraete
authors: Savvas N
authors: SN
authors: Savvides
authors: Bjorn
authors: B
authors: Vergauwen
mesh_terms: D055162:Biocatalysis; D003460:Crystallization; D008958:Models, Molecular; D010088:Oxidoreductases / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D041261:Vibrio vulnificus / Q000201:enzymology*
chemical_list: D010088:Oxidoreductases