In a standard assay for LDH activity we used a phosphate buffer, pyruvate, and NADH with...
In a standard assay for LDH activity we used a phosphate buffer, pyruvate, and NADH with the LDH enzyme. I don't understand what happens when we replace the NADH with NADPH and why does LDH have a preference for NADH over NADPH
Homework Answers
Preference of LDH for NADH is because specific interactions of the enzyme with NADH are just optimum. The only structural difference between NADH and NADPH is a free phosphate group which is absent in NADH. This extra phosphate group somehow destabilizes the interactions with the enzyme.
To completely understand this, you need to look at the crystal structure of LDH and LDH bound to NADH. In LDH, there is an aspartate(negatively charged) group at position 22 which interacts with a negatively charged phosphate group in NADPH (Negative-Negative charges close together destablize the interaction), is responsible for this preference.
You may refer to this https://www.ncbi.nlm.nih.gov/pubmed/10556245 paper for details.
I hope this explanation was of some help.
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