Crystal structure at 2.5 A resolution of zinc-substituted copper amine oxidase of Hansenula polymorpha expressed in Escherichia coli.
Copper amine oxidases (CAOs) catalyze the two-electron oxidation of primary amines to aldehydes, utilizing molecular oxygen as a terminal electron acceptor. To accomplish this transformation, CAOs utilize two cofactors: a mononuclear copper, and a unique redox cofactor, 2,4,5-trihydroxyphenylalanine quinone (TPQ or TOPA quinone). TPQ is derived via posttranslational modification of a specific tyrosine residue within the protein itself. In this study, the structure of an amine oxidase from Hansenula polymorpha has been solved to 2.5 A resolution, in which the precursor tyrosine is unprocessed to TPQ, and the copper site is occupied by zinc. Significantly, the precursor tyrosine directly ligands the metal, thus providing the closest analogue to date of an intermediate in TPQ production. Besides this result, the rearrangement of other active site residues (relative to the mature enzyme) proposed to be involved in the binding of molecular oxygen may shed light on how CAOs efficiently use their active site to carry out both cofactor formation and catalysis.
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Z
Z
Chen
B
B
Schwartz
N K
NK
Williams
R
R
Li
J P
JP
Klinman
F S
FS
Mathews
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry* / Q000235:genetics; D020134:Catalytic Domain; D003300:Copper / Q000737:chemistry*; D018360:Crystallography, X-Ray; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives / Q000096:biosynthesis; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010843:Pichia / Q000201:enzymology*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry; D014443:Tyrosine / Q000378:metabolism; D015032:Zinc / Q000737:chemistry*
D011994:Recombinant Proteins; D014443:Tyrosine; D004295:Dihydroxyphenylalanine; C067626:6-hydroxydopa quinone; D003300:Copper; D006631:Amine Oxidase (Copper-Containing); D015032:Zinc