11. The protein Gbg is involved in regulating a diverse array of downstream effe
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11. The protein Gbg is involved in regulating a diverse array of downstream effector mdtice where the authors designed channels, ion channels, adenylyl cyclase and lecules including calcium the authors molecules. You are now CAMP. Recently you discover a journal a peptide (that they called SIGK) that binds to Gbg and is able to selectively inhibit adenylyl cyclase but has no effect on ion-channels. With the help of a between SIGK and Gbg. After careful consideration you synthesize a total of 90 of these smalt thi left with the task of confirming whether these small molecules are capable of mimicking SIGK. In anticipation you obtain chemist you computationally design a series of small molecules that mimi the binding do a competition experiment test for binding of each individual small molecule to Gbg A. What simple experimental technique will you use to assay these 90 small molecules for to binding to Gbg (1 point) B. What additional reagents are needed other than the ones provided (1 point C. What would be appropriate positive and negative controls for interpreting the results (2 points) D. In a flow diagram sketch out the experimental steps needed to complete the experiment (2 points) E. After completing the binding study, you identify 2 candidate small molecules (M119 and M201) that are competing with SIGK peptide for binding to Gbg. You decide to test M119 and M201 efficacy in a mouse model of altered adenylyl cyclase activity. ADCY5 mice have hyperactive Gbg activity leading to increased adenyly cyclase expression in the gut. Your experiment consists of 4 cohorts of mice. After the completion of the experiment, the mice are sacrificed and sections from the gut are obtained and stained for active adenylyl cyclase. based on the figure provided red roasde he red arrows show cells with adenylyl cyclase expression. Answer the questions ) Comment on the effect of M201 and M119 compared to WT (2 points) b) Which drug would be a better candidate for a clinical trial and why? (2 points WT ADCY5+M201 WT - mice with normal expressic of adenylyl cyclase ADCY5 - mice with increased expression of adenylyl cyclase ABcyS ADCY5+M201 - ADCY5 mice treated with M201 ADCY5+M119 ADCY5+M119- ADCY5 mice treated with M119 iaura 1. Adanulul aulsen etainina of aut cactions from different miceExplanation / Answer
11.A.
Ligand receptor based assays could be conducted or a dye based assays could be conducted to find out the Gbg binding.
B) a flurocent dye is required for the experiment. Microarry analyzer and chip is also required to read the data.
C) a normal protein with the dye could be used as positive control and the experimental protein with the dye could be used as negative control.
C) array design - collect source peptide and organise them
Chip fabrication - use microfluid technology
Chip scanning - binding ligand receptor sites could be detected using the scanner
Data collection and processing - array pro analyzer and online tool which needs to collected to the computer and analyze the data.
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