RESUMO
CO dehydrogenase (EC 1.2.99.2) catalyzes the oxidation of CO according to the following equation: CO + H2O-->CO2 + 2 e- + 2 H+. It is a selenium-containing molybdo-iron-sulfur-flavoenzyme, which has been crystallized and structurally characterized in its oxidized state from the aerobic CO utilizing bacteria Oligotropha carboxidovorans and Hydrogenophaga pseudoflava. Both CO dehydrogenase structures show only minor differences, and the enzymes are dimers of two heterotrimers. Each heterotrimer is composed of a molybdoprotein, a flavoprotein, and an iron-sulfur protein. CO oxidation takes place at the molybdoprotein which contains a 1:1 mononuclear complex of molybdopterin-cytosine dinucleotide and a Mo-ion, along with a catalytically essential S-selanylcysteine. The latter is appropriately positioned in the SeMo-active site by a unique VAYRCSFR active site loop. In H. pseudoflava the arginine preceeding the cysteine in the active site loop is modified to a Cgamma-hydroxy arginine residue which has no obvious function. The substituents in the first coordination sphere of the Mo-ion are the enedithiolate sulfur atoms of the molybdopterin-cytosine dinucleotide, two oxo- and a sulfido-group. Extended X-ray absorption fine structure spectroscopy (EXAFS), along with the crystal structure of CO dehydrogenase (23.2 U mg(-1)) at 1.85 A resolution, have identified a sulfur atom at 2.3 A from the Mo-ion. The sulfur reacts with cyanide yielding thiocyanate. The corresponding inactive desulfo-CO dehydrogenase shows a typical desulfo inhibited-type of Mo-electron paramagnetic resonance (EPR) spectrum. Structural changes at the SeMo-site during catalysis are suggested by the Mo to Se distance of 3.7 A and the Mo-S-Se angle of 113 degrees in the oxidized enzyme which increase to 4.1 A, and 121 degrees, respectively, in the reduced enzyme. The intramolecular electron transport chain in CO dehydrogenase involves the following prosthetic groups and minimal distances: CO-->[Mo of the molybdenum cofactor] - 14.6 A - [2Fe-2S]I - 12.4 A - [2Fe-2S]II - 8.7 A - [FAD].
Assuntos
Aldeído Oxirredutases/metabolismo , Ferro/fisiologia , Molibdênio/fisiologia , Complexos Multienzimáticos/metabolismo , Selênio/fisiologia , Aldeído Oxirredutases/química , Animais , Catálise , Humanos , Complexos Multienzimáticos/química , Estrutura Secundária de ProteínaRESUMO
Millimeter-wave spectra of HSiF(3) and DSiF(3) in the v(3) = 1 excited state have been measured from 100 to 490 GHz. Infrared spectra have been recorded in the nu(3) regions, nu(0) 424.0301 and 420.9320 cm(-1) in HSiF(3) and DSiF(3), respectively, with a resolution of 2.4 x 10(-3) cm(-1). Since in both species the parameters alpha(B)(3) and alpha(C)(3) have very similar values, no K structure could be resolved in the (Q)P and (Q)R clusters for low-to-medium K values. For high J the effect of the ground state D(JK) term more and more dominates and spreads the J clusters into opposite directions such that medium-to-high K components, particularly those with K = 3p, are resolved. Rotational and infrared data have been fitted together using a model up to sextic centrifugal distortion constants. No perturbations were indicated. Hot bands (nu(3) + nnu(6))-nnu(6) with n = 1, 2, and 3 have been detected and analyzed. Copyright 2000 Academic Press.
RESUMO
The present paper deals with the analysis of the microwave, millimeter-wave, and infrared spectra of (28)SiHF(3) in its ground, v(6) = 1 and v(4) = 1 excited states. The former was observed up to 1055 GHz leading to the determination of one octic centrifugal distortion constant, L(J) = -0.0749(55) µHz. Furthermore the