Covalent binding of an NAD+ analogue to horse liver alcohol dehydrogenase in a ternary complex with pyrazole.
European journal of biochemistry
confidence
Key findings
NAD+ analogue covalently bound to horse liver alcohol dehydrogenase in ternary complex with pyrazole; acts as prosthetic group, reduced/reoxidized by coupled substrate reaction.
View source on PubMed (PMID 3665930) ↗
- Population
- In vitro (horse liver alcohol dehydrogenase)
- Dosing
- N6-[N-(8-amino-3,6-dioxaoctyl)carbamoylmethyl]-NAD+ analogue
- Route
- in vitro
- Blinding
- not_reported
- Controls
- none
- Drug class
- coenzyme
Full abstract
Examination of the model of the fixation site of the adenosine phosphate part of NAD+ on horse liver alcohol dehydrogenase led us to synthesize a NAD+ analogue N6-[N-(8-amino-3,6-dioxaoctyl)carbamoylmethyl]-NAD+ in order to alkylate the carboxylic acid group of Asp-273 and to convert the normally dissociable coenzyme into a permanently bound prosthetic group. This NAD+ analogue is coupled to the horse liver alcohol dehydrogenase in the ternary complex formed with pyrazole. In these conditions the degree of fixation varies between 0.4 and 0.58 coenzyme molecule/enzyme subunit molecule. The N6-[N-(8-amino-3,6-dioxaoctyl)carbamoylmethyl]NAD+ acts as a true prosthetic group which can be reduced and reoxidized by a coupled substrate reaction and the internal activity of this holoenzyme corresponds to the amount of analogue incorporated.