Biochemistry
Biochemistry
9th Edition
ISBN: 9781319114671
Author: Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher: W. H. Freeman
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Question

Why is phospholipase c is the target on this western blot explain in detail 

phospho-CREB ==== !! === phospho-ERK1/phospho-ERK2 phospho-MEK phospho-PKC Total GAPDH You should assume (i) that any phosphorylation event occurs as the results of the action of an upstream enzyme and (ii) that protein phosphorylation causes the activation of that protein. What is the likely molecular target of Drug X? Select all answers that apply Protein Kinase C МЕК Adenylyl cyclase Phospholipase C Protein Kinase A None of the above O OO S OO O
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Transcribed Image Text:phospho-CREB ==== !! === phospho-ERK1/phospho-ERK2 phospho-MEK phospho-PKC Total GAPDH You should assume (i) that any phosphorylation event occurs as the results of the action of an upstream enzyme and (ii) that protein phosphorylation causes the activation of that protein. What is the likely molecular target of Drug X? Select all answers that apply Protein Kinase C МЕК Adenylyl cyclase Phospholipase C Protein Kinase A None of the above O OO S OO O
Data from an experiment is shown in the figure below. In the experiment, murine cells were treated with a specific a ligand that activates receptor R. In some cases, the cells were exposed to one of two drugs (X or Y) as well as the ligand or were left untreated (UT). After 30 minutes of treatment, the cells were lysed with a detergent-based buffer to release the soluble membrane, cyosolic and nuclear proteins. Samples from each cell extract were run (in duplicate) by SDS-PAGE (SDS-polyacrylamide gel electrophoresis) in order to separate the proteins by molecular mass (size). The separated proteins were then transferred to nitrocellulose membrane which was then probed with different antibodies in a western blotting procedure to detect specific phosphorylated proteins or total proteins. If present in the cell extract, these proteins appear as a dark band in the relevant western blot image within the figure. Drug X Drug Y + UT Ligand Ligand Ligand Western blotting antibodies phospho-CREB phospho-ERK1/phospho-ERK2 phospho-MEK phospho-PKC Total GAPDH You should assume (i) that any phosphorylation event occurs as the results of the action of an upstream enzyme and (ii) that protein phosphorylation causes the activation of that protein.
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Transcribed Image Text:Data from an experiment is shown in the figure below. In the experiment, murine cells were treated with a specific a ligand that activates receptor R. In some cases, the cells were exposed to one of two drugs (X or Y) as well as the ligand or were left untreated (UT). After 30 minutes of treatment, the cells were lysed with a detergent-based buffer to release the soluble membrane, cyosolic and nuclear proteins. Samples from each cell extract were run (in duplicate) by SDS-PAGE (SDS-polyacrylamide gel electrophoresis) in order to separate the proteins by molecular mass (size). The separated proteins were then transferred to nitrocellulose membrane which was then probed with different antibodies in a western blotting procedure to detect specific phosphorylated proteins or total proteins. If present in the cell extract, these proteins appear as a dark band in the relevant western blot image within the figure. Drug X Drug Y + UT Ligand Ligand Ligand Western blotting antibodies phospho-CREB phospho-ERK1/phospho-ERK2 phospho-MEK phospho-PKC Total GAPDH You should assume (i) that any phosphorylation event occurs as the results of the action of an upstream enzyme and (ii) that protein phosphorylation causes the activation of that protein.
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