Question

A graduate student studying a rare green armadillo adds radiolabeled B-hydroxybutyrate so heart cells from the armadillo and no radiolabeled acetyl CoA is detected. In a separate experiment the concentrations of NAD and NADH in these cells was found to b normal. In another experiment radiolabeled acetoacetyl-CoA is added to these cells and radiolabeled acetyl CoA is detected. Explain what is wrong with this poor armadillo. Propose two experiments to test your hypothesis.

Answer 1

When radiolabeled beta - hydroxybutyrate injected to heart cell them - No NAD, NADN level change - No Radiolabeled Acetyl CoA But when Acetoacetyl CoA Radiolabeled injected then Radiolabeled Acetyl CoA defected So, we can say 1 Hypothesize that these may be defect in beta - hydroxybutyrate dehydrogenase enzyme. Experiment 1: - Gere for beta - hydroxybutyrate dehydrogenas enzyme might got muttased So, we can check Hypothesis by sequence of this enzyme gene from this poor armadillo of Normal Armadillo, with using proper standard protocol for sequencing by sangeis method. Then compose the sequence tells us about mutation in Gene, of enzyme is damaged at DNA level Experiment 2: - Checking of Enzyme activity at proseim level. By Enzyme assay method. Enzyme assay will be done for beta - hydroxybutyrate Dehydrogenase using its substrate as beta - hydroxybutyrate rate and essential cofactors as - NAD^+ then allow to assay & treatment using standard protocol. Finally check level of NAD^+ and NADN^+ Absorbance by spectrophotometer rightarrow If reaction will occurs than NADN^+