Introduction

In this laboratory you will perform simulations of biochemistry experiments designed to study the reactions of aerobic cellular respiration as it occurs in vitro using isolated preparations of mitochondria. By changing reaction conditions through adding substrates and metabolic inhibitors you will simulate some of the experiments that were used to determine the metabolic reactions and the sequence of the reactions that occur during the Krebs cycle and the electron transport chain.

Objectives

The purpose of this laboratory is to:

• Demonstrate how studying the effects of intermediates in a reaction can be used to determine the steps in a metabolic pathway and the sequence of each enzymatic step in the pathway.
  
• Demonstrate how changes in substrates, products, and other reaction parameters can effect the reactions involved in the Krebs cycle, electron transport chain, and oxidative phosphorylation.
  
• Simulate the effects of substances that inhibit the reactions of aerobic cellular respiration.

Before You Begin: Prerequisites

Before beginning MitochondriaLab you should be familiar with the following concepts:

• The importance and functions of enzymes as biological catalysts, basic principles of metabolic pathways, and mechanisms involved in regulating the catalytic activity of an enzyme.
• Structure and function of the mitochondrion.
• The reactions of aerobic cellular respiration to include glycolysis, the Krebs cycle, electron transport chain, and oxidative phosphorylation. Be able to describe the primary substrates required, reactions involved, and products generated by each of these reactions.
• The enzymatic steps involved in the Krebs cycle, and the organic acids produced as intermediates during the Krebs cycle.
• Feedback mechanisms involved in the control of cell respiration.