May 30- June 6 Establishing Controls

Stability of Relevant Molecules

For the first 2 weeks of summer research, the goal has been to lay the groundwork for the rest of the summer. The amount of time certain molecules can sit out on the lab top at room temperature before they degrade and become unusable had to be established. Then the effect of freezing and thawing an enzyme multiple times on its activity level was investigated. These things had to happen before the real research could begin. This took several days of control experiments.

Measurement of Pyruvate Kinase Activity
Next, the best way to measure the activity of the enzyme under study, pyruvate kinase, had to be determined. Initially, a different enzyme, lactate dehydrogenase, was used to measure the activity of pyruvate kinase. When this is done, the enzyme of interest makes its product. Then the measurement enzyme reacts with that product to make something measurable. This means that a reliable way to stop pyruvate kinase was needed. The use of heat and metal ions was used and both were found to be effective. The use of heat was preferable because of the lack of lingering ions.
At the very end of the first week and the beginning of the second a simple reaction was found. This reaction does not use enzymes, but still makes a measurable product from the product of pyruvate kinase. Then, the next few days were spent fine tuning the reaction and establishing what to expect from the reaction so it could be properly used.