NAD+-analogue, where the nicotinamide ribosyl moiety has been replaced by the nicotinamide (2-hydroxyethoxy)methyl moiety. The chemical properties are comparable to those of beta-NAD+ with a redox potential of -324 mV and a 341 nm lambdamax for the reduced form. The stereochemistry of the hydride transfer in the oxidation of n-butanol is identical to that for the reaction with beta-NAD+. There is no detectable reduction of acycloNAD+ by secondary alcohols although these alcohols serve as competitive inhibitors. AcycloNAD+ converts horse liver ADH from a broad spectrum alcohol dehydrogenase, capable of utilizing either primary or secondary alcohols, into an exclusively primary alcohol dehydrogenase
the binding of NAD+ is kinetically limited by a unimolecular isomerization (corresponding to the conformational change) that is controlled by deprotonation of the catalytic zinc-water to produce a negatively-charged zinc-hydroxide, which can attract the positively-charged nicotinamide ring
insertion of an RTX domain from the adenylate cyclase of Bordetella pertussis into a loop near the catalytic active site of the thermostable alcohol dehydrogenase D from Pyrococcus furiosus. The resultant chimera, beta-AdhD, gains the calcium-binding ability of the beta-roll, retains the thermostable activity of AdhD, and exhibits reduced overall alcohol dehydrogenase activity. The addition of calcium to beta-AdhD preferentially inhibits NAD+-dependent activity in comparison to NADP+-dependent activity. Calcium is a competitive inhibitor of AdhD, and the addition of the RTX domain introduces calcium-dependent noncompetitive inhibition to beta-AdhD affecting NAD+-dependent activity
the cofactor NADH can be recycled with D-glucose dehydrogenase/D-glucose system or in a coupled substrate approach using isopropanol as the hydrogen donor
insertion of an RTX domain from the adenylate cyclase of Bordetella pertussis into a loop near the catalytic active site of the thermostable alcohol dehydrogenase D from Pyrococcus furiosus. The resultant chimera, beta-AdhD, gains the calcium-binding ability of the beta-roll, retains the thermostable activity of AdhD, and exhibits reduced overall alcohol dehydrogenase activity. The addition of calcium to beta-AdhD preferentially inhibits NAD+-dependent activity in comparison to NADP+-dependent activity. Calcium is a competitive inhibitor of AdhD, and the addition of the RTX domain introduces calcium-dependent noncompetitive inhibition to beta-AdhD affecting NAD+-dependent activity
experiments with the highly active double mutant K249G/H255R using NADPH as a cofactor demonstrate an unprecedented transient behavior where the binding mechanism appears to be dependent on cofactor concentration
the purified ADHES77 prefers to use NAD+ as an electron acceptor as compared to NADP+. In the specific activity of ethanol dehydrogenase, the catalytic efficiency for NAD+ is about 56.7fold greater than for NADP+. Similarly, in the catalytic reduction of acetaldehyde, the ADHES77 preferentially utilizes NADH as an electron donor, exhibiting approximately 12.5fold higher activity, than for NADPH. The results suggest that the ADHES77 prefers to use NAD(H) as redox partners as compared to NADP(H)
LC-MS/MS analysis shows that Cys47 and Cys243 can make a stable disulfide bond with glutathione, suggesting redox sensitivity of these residues. Binding of ADH with its cofactors may limit availability of Cys residues to redox modifications
LC-MS/MS analysis shows that Cys47 and Cys243 can make a stable disulfide bond with glutathione, suggesting redox sensitivity of these residues. Binding of ADH with its cofactors may limit availability of Cys residues to redox modifications
the apoenzyme has an open conformation where the cleft between the coenzyme and catalytic domains allows coenzyme to bind. This structure is similar to those of the apoenzyme forms determined by X-ray crystallography (PDB ID 4W6Z). The complex made with NADH also has an open conformation and the inverted zinc coordination, and NADH is bound to the coenzyme binding domain in a position similar to that found for NAD+ in the B chain subunit in the X-ray structure (PDB ID 5ENV). In contrast, the structure for the sample of the enzyme made with NADH and pyrazole has the closed conformation with NADH bound in the active site and the classical coordination of the zinc with a water