How does a competitive binding assay determine the affinity for an agonist? Specifically, how is the competition measured and the number of receptors/ligands counted?
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How does a competitive binding assay determine the affinity for an agonist? Specifically, how is the competition measured and the number of receptors/ligands counted?
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- In a radiolabeled ligand assay to determine the binding affinity for an agonist, different concentrations of agonist are placed in test tubes with both a set number of receptors and radiolabeled ligand. Filters separate the bound and free radioligand, and a scintillation counter measures the number of bound radioligand. But how is the binding affinity of the agonist determined from all this?In a radiolabeled ligand assay to determine the binding affinity for an agonist, how is the amount of bound radioligand measured after filtration?What is signal transduction? Illustrate and describe the molecular events in signal transduction pathways involving 1) G-protein-coupled receptors and 2) enzyme linked receptors.
- How do specific and nonspecific binding factor into calculations for a competitive binding assay to find the affinity of an agonist? For example, are they calculated before or after the assay, and does it affect the binding isotherm curve and how it is calculated?What is the importance of understanding the interactions between the receptor binding domain of the spike protein and ACE 2? List four of the interactions (hydrogen bonds) between the spike protein and the receptor (ACE 2)?My boss has given me the purified extracellular portion of the Notch receptor. As Delta/Notch signaling occurs between cells, I know that both the Notch receptor and and Delta are most likely "comfortable" in the environment of the blood, not the cytosol, nucleus, mitochondria, etc... I want to purify Delta, the signal. What would be true of the most specific purification technique I could choose, i.e. the technique that will result in the fewest proteins in the fraction containing Delta? Do not worry about whether the protein works after fractionation yet. Pick ALL that apply. I would use ion exchange chromatography. I would use size exclusion chromatography I would use affinity chromatography I would use a water based mobile phase of 7.4 I would use an acetonitrile mobile phase with a pH of 7.4 I would use a water based mobile phase of pH 7.2 I would use an acetonitrile phase of 7.2 I…
- Using molecular cloning, a hybrid receptoer is produced that consists of the extracellular domain of the B-adrenergic receptor (B-AR) and the transmembrane and intracellular domains of the EGF receptor (EGFR). Question: What is the stoichiometric ratio of epinephrine to the hybrid receptor (B-AR-EGFR)? 1:1 O 1:4 O 1:2 2:1 O O OWhat are all the different ways that the g-protein coupled receptor pathway can be targeted by drugs? Please specify the mechanisms and locations with examples.Describe the signal transduction pathway for Two or Three of the following: a) ß-adrenergic (epinephrine) receptor b) Histamine receptor (hint: this is a Go type receptor) c) Growth hormone receptor d) Epidermal growth factor receptor e) Insulin receptor Make sure to cover all the principal steps within molecular circuits in the specifically chosen signal transduction pathway. Also, be clear on which exact receptor and pathway you are describing. You can support your answer with a drawing, but do not forget to describe it.
- Sort the following steps of GPCR signal propagation (you may not use all the steps): A) Dephosphorylation of GPCR B) G protein dissociation to adenylate cyclase and GTP binds to the G protein C) Ligand binding to GPCR D) G protein dissociation from GPCR and GDP binds to the G protein E) Synthesis of CAMP F) Activation of PKA G) G protein association to adenylate cyclase H) G protein dissociation from GPCR and GTP binds to the G protein O F, C, H, F, B OC OB, D, A, E, H O C, H, G, E, F OA, G, H, B, EInhibitors of acetylcholinesterase, such as edrophonium, are used to treat Alzheimer’s disease. The substrate for acetylcholinesterase is acetylcholine. Structures are attached. What kind of inhibitor is edrophonium? Explain. Can inhibition by edrophonium be overcome in vitro by increasing the substrate concentration? Explain. Does this inhibitor bind reversibly or irreversibly to the enzyme? Explain.Control wells in the dopamine assay normally look like this: dopamine HVA Legend: = high (dopamine] = low (or no) (dopamine] neural cells If the researchers accidentally added neural cells to all of the wells, what would the control wells look like?