Lab 5 Flashcards
Objectives
- Relate general metabolic activity to specific enzymes in metabolic pathways.
-Use normalized enzyme activity to make quantitative comparisons between tissues.
-Explain the design of an indirect enzyme assay
-Explain the concept of competitive enzyme inhibition.
-Describe the role of positive and negative controls in an experiment.
Metabolism
The total of all the anabolic and catabolic reactions of a cell.
Tissues that perform many metabolic reactions require
High levels of ATP
Most ATP in cells is made
During oxidative phosphorylation in mitochondria
Oxidative phosphorylation is
Powered by the proton gradient created by the electron transport, which in turn is driven by NADH
NADH comes from
The citric acid cycle
Since much NADH comes from the citric acid cycle
Levels of citric acid enzymes can be used as a marker of metabolic rates of a tissue
First thing that I will be doing is
Measure the activity of a citric acid cycle enzyme in homogenates from three organs of B. Taurus and then use that value to figure out the metabolic activity of the three organs.
Oxidative phosphorylation Step 1
The citric acid cycle produces NADH and FADH2, which are oxidized by the electron transport chain in order to power movement of protons into the intermembrane space.
Oxidative phosphorylation Step 2
Protons then flow down their concentration gradient into the matrix through ATP synthase, powering ATP synthesis.
One of the enzymes in the citric acid cycle
Succinate dehydrogenase
Succinate dehydrogenase
Is Transported to the mitochondria after synthesis on ribosomes
FADH2 has a similar function to NADH
In that it donates electrons to the electron transport chain
oxidized DCIP
Blue
reduced DCIP
No Color
