Question
I know these are basically the same question but I just wanted to get clarification on what answers would be true or false and what other sorts of conclusions can be made based on the information given.
You take total cell lysate, and use immunoprecipitation to purify an enzyme of interest. The purified enzyme has HIGH specific activity, and shows a single band (56kD) on an SDS PAGE. Now, you heat the total cell lysate, and use immunoprecipitation to purify the enzyme of interest. The purified enzyme has LOW specific activity, and shows two bands (56kD and 122kD) on an SDS PAGE You then heat the total cell lysate, allow it to cool down, and use immunoprecipitation to purify the enzyme of interest. The purified enzyme has HIGH specific activity, and shows one band (56kD) on an SDS PAGE. Finally, you make total cell lysate from a mutant cell. When you use immunoprecipitation to purify the enzyme of interest, the purified enzyme has LOW specific activity, and shows a single band (56kD) on an SDS PAGE. When you heat the cell lysate from the mutant cells and immunoprecipitate the enzyme, the purified enzyme has LOW specific activity, and shows a single band (56kD) on an SDS PAGE. What is the BEST conclusion from this experiment?
Explanation / Answer
1. False
The antibody picked the E+HSP complex.
1 band for the pure enzyme and the other band for the complex.
2. True
The other protein could be a chaperone that helps in proper refolding of the protein.
3. True
The mutant protein could be ineffective in refolding as it shows less activity after heat-cooling cycle.
4. False
Chaperones positively affect protein folding.