Methyl-Coenzyme M Reductase Binding Experiments
Project Description
MULTIPLE TESTING
Contract Laboratory needed for experiments on binding assays to determine the Kd Values of several Methyl-Coenzyme M Reductase potential binders (inhibitors, not substrates) in an anaerobic chamber.
What we require are the Kd values of binding because they can differ significantly from the Ki values in this case. To obtain such values we need to have comparable experiments with the type of assay as described by this paper: T Wongnate et. al. The Reaction Mechanism of Methyl-Coenzyme M Reductase. J Biol Chem . 2015 Apr 10;290(15):9322-34. doi:10.1074/jbc.M115.636761.
Because of the fast oxidation of Ni+1 these assays should be performed in an anaerobic chamber. The techniques may be modified, but our end goal is to reproduce the results from the above mentioned paper and in particular this: Fluorescence and EPR results clearly demonstrate that either substrate can bind tightly to the enzyme in the absence of the other substrate, with the Kd value for the MCRCoB7SH complex(90 +/-22uM) being only 7?18-fold weaker than that for the MCR-methyl-SCoM complex (5?13uM)?.
However, I would also like to receive a quote for the same experiments but with Ni+2 (II) (silent) state, which will probably be more simple. In fact, if you suggest a faster and cheaper approach with the silent Ni+2 Methyl-Coenzyme M Reductase form/state we can start with this.
Project Information
Number:23-01943
