Crystallographic and spectroscopic studies of native, aminoquinol, and monovalent cation-bound forms of methylamine dehydrogenase from Methylobacterium extorquens AM1.
Various monovalent cations influence the enzymatic activity and the spectroscopic properties of methylamine dehydrogenase (MADH). Here, we report the structure determination of this tryptophan tryptophylquinone-containing enzyme from Methylobacterium extorquens AM1 by high resolution x-ray crystallography (1.75 A). This first MADH crystal structure at low ionic strength is compared with the high resolution structure of the related MADH from Paracoccus denitrificans recently reported. We also describe the first structures (at 1.95 to 2.15 A resolution) of an MADH in the substrate-reduced form and in the presence of trimethylamine and of cesium, two competitive inhibitors. Polarized absorption microspectrophotometry was performed on single crystals under various redox, pH, and salt conditions. The results show that the enzyme is catalytically active in the crystal and that the cations cause the same spectral perturbations as are observed in solution. These studies lead us to propose a model for the entrance and binding of the substrate in the active site.
Labesse
G
G
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Ferrari
D
D
Chen
Z W
ZW
Rossi
G L
GL
Kuusk
V
V
McIntire
W S
WS
Mathews
F S
FS
D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D002586:Cesium / Q000494:pharmacology; D018360:Crystallography, X-Ray; D045563:Indolequinones; D008744:Methylamines / Q000378:metabolism / Q000494:pharmacology; D008958:Models, Molecular; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry*; D011809:Quinones / Q000737:chemistry*; D013053:Spectrophotometry; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000737:chemistry
D001426:Bacterial Proteins; D045563:Indolequinones; D008744:Methylamines; D011809:Quinones; C070569:tryptophan tryptophylquinone; D002586:Cesium; D014364:Tryptophan; C027451:methylamine; C021567:methylamine dehydrogenase; D000587:Oxidoreductases Acting on CH-NH Group Donors; C023336:trimethylamine