4. You develop the covalently-acting irreversible kinase inhibitor ibrutinib shown on the left that reacts potently with a cysteine in the ATP binding pocket of the cancer-promoting kinase Bruton's Tyrosin Kinase (BTK). You would like to know whether ibrutinib engages and selectively targets BTK in vivo in the tumor of a mouse model of cancer. Describe experimentally how you would use activity-based protein profiling to assess whether ibrutinib inhibited BTK in vivo in the tumor and also determine how selectively ibrutinib engaged BTK compared to other kinases in the tumor? ( NH, H₂N- N ibrutinib OH OH OH OH activity-based probe for kinases that covalently modifies active-site lysine of all kinase ATP binding pocket W ta t C
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- I just read an abstract of the paper “Disulfide bond-disrupting agents activate the tumor necrosis family-related apoptosis-inducing ligand/death receptor 5 pathway” and noted that “DDAs and TRAIL synergize to kill cancer cells and are cytotoxic to HER2+ cancer cells with acquired resistance to the EGFR/HER2 tyrosine kinase inhibitor Lapatinib.” For the last sentence, I am not sure the meaning of the “acquired resistance to the EGFR/HER2 tyrosine kinase inhibitor Lapatinib”. Is the “acquired resistance ... to inhibitor” a good thing or bad thing, as far as the synergize effect of DDAs and TRAIL”? Hope that expert can help.Suppose that Protein J which is a hypothetical protein kinase receptor was determined to be related to the progression of cancer through its activation. It was also determined that the protein exists in the active and inactive forms. The said active form is highly similar to the Protein K's conformation. Ligands A, B, and C, which are lead inhibitor compounds, were optimized to inhibit Protein J. The affinities of the ligands are shown in the table. Kp values Active Protein J Inactive Protein J Protein K Ligand A 10 mM 20 nM 5 mM Ligand B 20 nM 10 mM 15 nM Ligand C 20 nM 15 nM 15 nM Question: a. Which of the ligands, based on the table, has the highest specificity in binding to the target Protein J?Suppose that Protein J which is a hypothetical protein kinase receptor was determined to be related to the progression of cancer through its activation. It was also determined that the protein exists in the active and inactive forms. The said active form is highly similar to the Protein K's conformation. Ligands A, B, and C, which are lead inhibitor compounds, were optimized to inhibit Protein J. The affinities of the ligands are shown in the table. Kp values Active Protein J Inactive Protein J Protein K Ligand A 10 mM 20 nM 5 mM Ligand B 20 nM 10 mM 15 nM Ligand C 20 nM 15 nM 15 nM Question: a. Which of the ligands, based on the table, may be expected to be the most potent or have the highest activity against cancer? Explain. b. Which of the ligands, based on the table, may be expected to be least toxic to normal cells? Explain.
- Suppose that Protein J which is a hypothetical protein kinase receptor was determined to be related to the progression of cancer through its activation. It was also determined that the protein exists in the active and inactive forms. The said active form is highly similar to the Protein K's conformation. Ligands A, B, and C, which are lead inhibitor compounds, were optimized to inhibit Protein J. The affinities of the ligands are shown in the table. Kp values Active Protein J Inactive Protein J Protein K Ligand A 10 mM 20 nM 5 mM Ligand B 20 nM 10 mM 15 nM Ligand C 20 nM 15 nM 15 nM Question: Describe the relative binding affinities of Ligand A to Protein K and to the active and inactive forms of Protein J. Determine which will Ligand A bind with the highest, medium, and lowest affinity.1.Describe in detail the signal transduction pathway that leads to activation of either PKA, Kinase or PC. 2. Describe in detail the signal transduction pathway that leads to activation of MAPKinase or Akt/PKB. 3.Describe the similarities and differences in the structures of GPCRs specific for various ligands including the extracellular , transmembrane , and intracellular domains.10. Explain how scaffold proteins help the efficiency of the AMP kinase cascade. Why is it important that the interaction of the scaffold with the MAP kinases is NOT a very stable interaction?
- The PYK gene codes for the expression of pyruvate kinase, which is one of the enzymestargeted for anti-cancer drug design. You have identified an RNAi that targets the mRNAof PYK gene. To study the effect of the RNAi towards pyruvate kinase, the respected RNAiis expressed in Saccharomyces cerevisiae. The level of pyruvate kinase can be detectedwith a fluorescent antibody.(a). Predict the result that you will obtain in recombinant S. cerevisiae that expresses therespected RNAi.(b). Compare the result in Q3a(i) with the wild-type S. cerevisiae.Although numerous treatments are available for advanced hormone receptor- positive (HR+) and human epidermal growth factor receptor 2 negative (HER2-) breast cancers, they are rarely curable. Drugs such as palbociclib that inhibit cyclin-dependent kinase 4 and 6 (CDK4/6) are rapidly altering this therapeutic environment. Discuss the function of CDK4 and CDK6 in promoting cell growth as well as the overall effects of their inhibition by palbociclib on the progression of the cell cycle.Drug X shown below is a kinase inhibitor used to treat multiple types of breast cancer. Breast cancer cells are treated with the drug and after 8 hr are lysed and the presence of various proteins is shown by a band on a western blot. The presence of the phosphorylated form of the protein is shown by the presence of a band when probed with antibodies that recognize the phosphorylated form of protein and are indicated by a p- in front of protein name (e.g. p-AKT). Based on this information, which kinase is likely the target and is inhibited by Justify your answer in 3-4 sentences. Drug X. F FF N. -N S IZ H₂N `N No Drug (+) Drug p-AKT AKT p-GSK3B GSK3B p-p70S6K p70S6K
- You are given a project to characterize the effect of mutation on two newly discovered G protein coupled-receptors, GPCR-UCI and GPCR-UC2, both of which bind to the same ligand. For the cell signalling assay, you have the UMR-106 cell line that expresses both GPCR-UC1, CPCR- UC2, the corresponding G proteins, and adenyl-cyclase. A basal adenylyl cyclase activity, and thus, a baseline cAMP concentration, is detected in the cell line. (i) You found that, upon binding to the ligand, the mutation in GPCR-UC1 results in increased CAMP level but the mutation in GPCR-UC2 results in a low cAMP concentration. Based on these results, identify the subclass of G protein isoforms that the GPCRs activate. Explain your answer. (ii) If the mutation affects the G protein from GDP release, how would this affect the CAMP production after ligand binding? Explain your answer.Doxorubicin is a chemotherapeutic drug with an IC50 in MCF7 breast cancer cells of 0.01 microM. Assuming the cytotoxicity of doxorubicin follows 'classic' receptor theory what percentage of the cells will be viable following treatment of the cells with a concentration of doxorubicin of 0.01 microM? O 10% O 0.01% O 90% 50% 99.99%Which ONE of these statements is the most accurate definition of the mode of action of imatinib? Select one: A.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks phosphatase activity by competing with adenosine triphosphate (ATP) binding B.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks tyrosine kinase activity by competing with adenosine triphosphate (ATP) binding C.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks tyrosine kinase activity by interaction with the enzyme site D.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks phosphatase activity by interaction with the enzyme site