Crystallographic study on the interaction of L-lactate oxidase with pyruvate at 1.9 Angstrom resolution.
L-Lactate oxidase (LOX) from Aerococcus viridans catalyzes the oxidation of L-lactate to pyruvate by the molecular oxygen and belongs to a large family of 2-hydroxy acid-dependent flavoenzymes. To investigate the interaction of LOX with pyruvate in structural details and understand the chemical mechanism of flavin-dependent L-lactate dehydrogenation, the LOX-pyruvate complex was crystallized and the crystal structure of the complex has been solved at a resolution of 1.90 Angstrom. One pyruvate molecule bound to the active site and located near N5 position of FMN for subunits, A, B, and D in the asymmetric unit, were identified. The pyruvate molecule is stabilized by the interaction of its carboxylate group with the side-chain atoms of Tyr40, Arg181, His265, and Arg268, and of its keto-oxygen atom with the side-chain atoms of Tyr146, Tyr215, and His265. The alpha-carbon of pyruvate is found to be 3.13 Angstrom from the N5 atom of FMN at an angle of 105.4 degrees from the flavin N5-N10 axis.
Li
Shu Jie
SJ
Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
Umena
Yasufumi
Y
Yorita
Kazuko
K
Matsuoka
Takeshi
T
Kita
Akiko
A
Fukui
Kiyoshi
K
Morimoto
Yukio
Y
D001665:Binding Sites; D003198:Computer Simulation; D003461:Crystallography / Q000379:methods*; D004789:Enzyme Activation; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000648:ultrastructure*; D008956:Models, Chemical; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D019289:Pyruvic Acid / Q000737:chemistry*; D012680:Sensitivity and Specificity
D019289:Pyruvic Acid; D006899:Mixed Function Oxygenases; C021497:lactate 2-monooxygenase