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PHTX 537 EXAMINATION 1 2002 DATE: Thursday, September 12th FROM: Edward JN Ishac, Course Director NOTE: This exam is scheduled to last for 2 HOURS. Apportion your time to approximate 1 minute per point value. Please write the last 4 digits of your social security number on each page of the exam. Also please answer the questions from each lecturer on separate pages from those of the others. Finally, before answering the following questions, read and analyze them carefully to identify the information being requested. When you have done this, write your response, using the same organizational format as the question. Be sure to address each point raised in the question, but you are strongly discouraged from including extraneous information not asked for (even though you may be very eager to demonstrate your knowledge). There are FIVE exams scheduled for this course. The questions will be as they were in the past, i.e. multiple choice questions, short answer and/or essay type questions. Each exam will be worth 100 points, so figure on an 8-10 point question that should take 8-10 minutes to answer per lecture hour. There is no cumulative final exam, so all exams are weighted evenly in the final grade. GOOD LUCK PHTX 537 Receptor: Cell Signaling Fall Semester 2002 Dr. Selley Examination 1 55 points total Section 1: Short answer (12 points total, 2 pts each) 1. What is the rate-limiting step in G-protein activation? 2. Second messenger-activated kinases phosphorylate which two amino acid residues on proteins? 3. What family of G is most associated with stimulation of phospholipase C? 4. Phospholipase D (PLD) generates which second messenger in large amounts? 5. Soluble guanylyl cyclases are most strongly activated by what? 6. The G-gamma-like (GGL) domain of many large regulators of G-protein signaling (RGS) proteins, allows the RGS to heterodimerize with what? 2 Receptor: Cell Signaling Dr. Selley continued Section 2: Short answer (15 points total, 3 pts each) 7. What are two ways that regulators of G-protein signaling (RGS) proteins can attenuate G-protein signaling? 8. Ca++/calmodulin kinase (CaMK) type IV is maximally activated when which two conditions are present? 9. The major isoforms of protein kinase C (PKC) are activated by which two second messengers? 10. What are two ways by which adenylyl cyclase type I can be inhibited? 11. What are two factors that can stimulate epithelial nitric oxide synthase (eNOS)? 3 Receptor: Cell Signaling Dr. Selley continued Section 3: Upstream/downstream. For each of the following signaling components, please identify (12 points total, 3 pts each): a) what it does b) the immediate upstream activator c) a downstream target 12. cGMP-phosphodiesterase a. b. c. 13. Protein kinase A (PKA) a. b. c. 14. NMDA receptor a. b. c. 15. Inositol triphosphate (IP3) receptor a. b. c. 4 Receptor: Cell Signaling Dr. Selley continued Section 4: Short essay (16 points total) 16. Describe the basic structure, function and mechanism of action of the cytoplasmic steroid type receptors? (7 points) 17. Describe start to finish the process of homologous desensitization and downregulation of a GPCR by G-protein-coupled receptor kinase 2 (GRK2) (9 points). 5 Receptor Theory Dr. Martin 45 points total Question 1. (20 points) Please compare the theories of Clark, Stevenson and Paton as follows: A. State the basic premise of each theory B. What is the contribution of each theory C. Describe limitations of each theory D. Define agonist, partial, and antagonist according to each theory Question 2. (15 points). Please provide a brief description of how receptor affinity and receptor number are determined using binding assays. What are the criteria that are use to distinguish receptors from non-receptor binding sites. Question 3. (10 points) Discuss the ternary model that best describes the equilibrium between a ligand, receptor and G-protein at both resting and activated states. 6