Question

help please

PART III: This part has to be done on your own and at the end of your experiments you will have to collect your data to write a labreport. You have to apply a rigorous scientific method by proposing and testing a hypothesis, select variables and design your own controlled experiment to collect data The purpose is to study the effect of denaturation on proteins' function. To this end, you will have to pick three denaturing agents among those tested in PART II and study their effect on the function of one the 2 following proteins: amylase or invertase. Reminder Amylase is an enzyme that catalyzes the hydrolysis of starch into simple reducing sugars while invertase catalyzes the hydrolysis of sucrose into fructose and glucose (Lab 2 carbohydrates). Question 1. Following denaturation, how are you going to check if these enzymes are still functional or not? In other words, what tests can be done to verify these enzymes' functions? Question 2: What can you conclude about the experiments done in PART II? In other words, how are you going to select the denaturing agents to use in PART III? Give at least 2 arguments that justify your choice. Question 3: How can you check if at the end of your experiment your protein is not totally degraded? In other words, what test can you do to show that the protein is still present but not functional?

Answer 1

1.Protein profiling is to be done for identification of protein and its types. Methods such as SDS PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis ), Chromatography, NMR (Nuclear Magnetic Resonance) can help to study the nature and structure of proteins. Obviously, denatured and non denatured proteins will have different mass to charge ratio,structure etc.. so you can identify.The above mentioned methods are expensive methods, we can do it through economic methods like Denaturation studies, Voltammetry and spectroscopy studies.

2.Selection of denaturing agents can b done based on these method itself.You can run normal denatured protein and also the various denatured ones,then by comparing results you can identify which of the denaturing agents show the difference in protein structure compared to non-denatured control.Then select those agents that has the capacity to denature the proteins ,they can be used for next experiment.

3.In general any molecular assays like NMR, Chromatography, PAGE etc.. That are used to study protein structure can be used for denaturation comparison studies