1U7H

Structure and a Proposed Mechanism for Ornithine Cyclodeaminase from Pseudomonas putida

1U7H の概要

• エントリーDOI: 10.2210/pdb1u7h/pdb
• 分子名称: ornithine cyclodeaminase, SODIUM ION, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, ... (5 entities in total)
• 機能のキーワード: deaminase, binds nad+, binds l-ornithine, binds l-pro, helix bundle, beta barrel, rossmann fold, lyase
• 由来する生物種: Pseudomonas putida
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 79659.66

構造登録者

Alam, S.,Goodman, J.L.,Wang, S.,Ruzicka, F.J.,Frey, P.A.,Wedekind, J.E. (登録日: 2004-08-03, 公開日: 2004-11-09, 最終更新日: 2024-10-30)

主引用文献

Goodman, J.L.,Wang, S.,Alam, S.,Ruzicka, F.J.,Frey, P.A.,Wedekind, J.E.
Ornithine Cyclodeaminase: Structure, Mechanism of Action, and Implications for the u-Crystallin Family;
Biochemistry, 43:13883-13891, 2004

PubMed Abstract: Ornithine cyclodeaminase catalyzes the conversion of L-ornithine to L-proline by an NAD(+)-dependent hydride transfer reaction that culminates in ammonia elimination. Phylogenetic comparisons of amino acid sequences revealed that the enzyme belongs to the mu-crystallin protein family whose three-dimensional fold has not been reported. Here we describe the crystal structure of ornithine cyclodeaminase in complex with NADH, refined to 1.80 A resolution. The enzyme consists of a homodimeric fold whose subunits comprise two functional regions: (i) a novel substrate-binding domain whose antiparallel beta-strands form a 14-stranded barrel at the oligomeric interface and (ii) a canonical Rossmann fold that interacts with a single dinucleotide positioned for re hydride transfer. The adenosyl moiety of the cofactor resides in a solvent-exposed crevice on the protein surface and makes contact with a "domain-swapped"-like coil-helix module originating from the dyad-related molecule. Diffraction data were also collected to 1.60 A resolution on crystals grown in the presence of l-ornithine. The structure revealed that the substrate carboxyl group interacts with the side chains of Arg45, Lys69, and Arg112. In addition, the ammonia leaving group hydrogen bonds to the side chain of Asp228 and the site of hydride transfer is 3.8 A from C4 of the nicotinamide. The absence of an appropriately positioned water suggested that a previously proposed mechanism that calls for hydrolytic elimination of the imino intermediate must be reconsidered. A more parsimonious description of the chemical mechanism is proposed and discussed in relation to the structure and function of mu-crystallins.

PubMed: 15518536
DOI: 10.1021/bi048207i

実験手法

X-RAY DIFFRACTION (1.8 Å)