Ornithine decarboxylase promotes catalysis by binding the carboxylate in a buried pocket containing phenylalanine 397.

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Jackson LK, Brooks HB, Myers DP, Phillips MA

Ornithine decarboxylase promotes catalysis by binding the carboxylate in a buried pocket containing phenylalanine 397.

Biochemistry. 2003 Mar 18;42(10):2933-40.

PubMed ID

12627959 [ View in PubMed

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Abstract

Ornithine decarboxylase (ODC) is a pyridoxal 5'-phosphate (PLP) dependent enzyme that catalyzes the decarboxylation of l-Orn to putrescine, a rate-limiting step in the formation of polyamines. The X-ray crystal structures of ODC, complexed to several ligands, support a model where the substrate is oriented with the carboxyl-leaving group buried on the re face of the PLP cofactor. This binding site is composed of hydrophobic and electron-rich residues, in which Phe-397 is predicted to form a close contact. Mutation of Phe-397 to Ala reduces the steady-state rate of product formation by 150-fold. Moreover, single turnover analysis demonstrates that the rate of the decarboxylation step is decreased by 2100-fold, causing this step to replace product release as the rate-limiting step in the mutant enzyme. These data support the structural prediction that the carboxyl-leaving group is positioned to interact with Phe-397. Multiwavelength stopped-flow analysis of reaction intermediates suggests that a major product of the reaction with the mutant enzyme is pyridoximine 5'-phosphate (PMP), resulting from incorrect protonation of the decarboxylated intermediate at the C4' position. This finding was confirmed by HPLC analysis of the reaction products, demonstrating that Phe-397 also plays a role in maintaining the integrity of the reaction chemistry. The finding that the carboxylate-leaving group is oriented on the buried side of the PLP cofactor suggests that ODC facilitates decarboxylation by destabilizing the charged substrate carboxyl group in favor of an electrostatically more neutral transition state.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Pyridoxal phosphate	Ornithine decarboxylase	Protein	Humans	Unknown	Cofactor	Details