Dear All,
I'm planning an experiment to study the oxidation of NADH by a
flavoprotein at cryogenic temperatures to facilitate collection of X-
ray diffraction data.
In planning this experiment, I have seen a few obstacles that I am
looking for help in overcoming.
1. There are no structures in the PDB that are complexed with NADH or
NAD2H.
Has anyone ever attempted to solve a structure complexed with NADH or
NAD2H, especially at cryogenic temperatures, and if so, what are the
difficulties? Does NAD+ de-bind from the protein too fast to permit
data collection?
2. NADH oxidation typically takes less than a second by a flavoprotein
at room temperature.
Is there an NADH or NAD2H analog that has a much longer half time for
oxidation by a flavoprotein, for example tens of minutes, rather than
tenths of a second, and can this analog still be oxidized at cryogenic
temperatures, with a reasonable half time, of several hours or so?
Thanks! and all the best,
--Buz
