Structure of the thiamine- and flavin-dependent enzyme pyruvate oxidase

Pyruvate oxidase from Lactobacillus plantarum is a tetrameric enzyme that decarboxylates pyruvate, producing hydrogen peroxide and the energy-storage metabolite acetylphosphate. Structure determination at 2.1 angstroms showed that the cofactors thiamine pyrophosphate (TPP) and flavin adenine dinucleotide (FAD) are bound at the carboxyl termini of six-stranded parallel beta sheets. The pyrophosphate moiety of TPP is bound to a metal ion and to a beta alpha alpha beta unit corresponding to an established sequence fingerprint. The spatial arrangement of TPP and FAD suggests that the oxidation of the oxyethyl intermediate does not occur by hydride displacement but rather by a two-step transfer of two electrons.

[1]  M G Rossmann,et al.  Comparison of super-secondary structures in proteins. , 1973, Journal of molecular biology.

[2]  M. Karplus,et al.  Crystallographic R Factor Refinement by Molecular Dynamics , 1987, Science.

[3]  G. Schulz.  Binding of nucleotides by proteins , 1992, Current Biology.

[4]  W. Hol,et al.  Analysis of the active site of the flavoprotein p-hydroxybenzoate hydroxylase and some ideas with respect to its reaction mechanism. , 1990, Biochemistry.

[5]  G. Schneider,et al.  Three‐dimensional structure of apotransketolase flexible loops at the active site enable cofactor binding , 1992, FEBS letters.

[6]  A. Schellenberger.  Die Funktion der 4′‐Aminopyrimidin‐Komponente im Katalysemechanismus von Thiaminpyrophosphat‐Enzymen aus heutiger Sicht , 1990 .

[7]  R. Wierenga,et al.  INTERACTION OF PYROPHOSPHATE MOIETIES WITH ALPHA-HELIXES IN DINUCLEOTIDE BINDING-PROTEINS , 1985 .

[8]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[9]  G. Schneider,et al.  Three‐dimensional structure of transketolase, a thiamine diphosphate dependent enzyme, at 2.5 A resolution. , 1992, The EMBO journal.

[10]  G. Bricogne,et al.  Methods and programs for direct‐space exploitation of geometric redundancies , 1976 .

[11]  Jeremy B. A. Green.  Pyruvate decarboxylase is like acetolactate synthase (ILV2) and not like the pyruvate dehydrogenase E1 subunit , 1989, FEBS letters.

[12]  P. Karplus,et al.  Refined structure of glutathione reductase at 1.54 A resolution. , 1987, Journal of molecular biology.

[13]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[14]  R. Perham,et al.  A common structural motif in thiamin pyrophosphate‐binding enzymes , 1989, FEBS letters.

[15]  R. Gennis,et al.  Role of the divalent metal cation in the pyruvate oxidase reaction. , 1982, The Journal of biological chemistry.