The Pharmacology of Functional, Biochemical, and Recombinant Receptor Systems

by Kenakin, Terrence P.; Angus, J. A.; Black, J. W. (CON); Barr, A. J. (CON); Bond, Robert A. (CON); Bouvier, M. (CON)

• ISBN13: 9783540661245
• ISBN10: 3540661247
• Format: Hardcover
• Copyright: 2000-09-01
• Publisher: Springer Nature

Summary

The technology of drug testing is rapidly advancing. This book brings together world renowned experts in the fields of pharmacology and physiology united with the common theme of describing methods, theoretical concepts and in vitro systems used to study drug receptor mechanisms and the action of drugs on receptors. The first section deals with the physiology and pharmacology of integrated natural systems, the second with the new theoretical ideas used to model receptor systems (i.e. the nature of efficacy, receptor activation) and the third with the new in vitro systems available for the study of receptors and drugs. The latest ideas regarding receptor theory are critically evaluated and presented to the reader.

Table of Contents

Introduction: Bioassays - Past Uses and Future Potential	1	(15)
J.W. Black
Section I: Classical Pharmacology and Isolated Tissue Systems
Human Vascular Receptors in Disease: Pharmacodynamic Analyses in Isolated Tissue	15	(36)
J.A. Angus
Introduction	15	(1)
Receptors Mediating Coronary Artery Contraction: Role in Variant Angina	16	(12)
Large Coronary Arteries	16	(1)
Ergometrine	16	(1)
5-HT and Endothelium-Derived Relaxing Factor	16	(5)
Small Resistance Arteries	21	(6)
Summary and Future Work	27	(1)
Vascular Reactivity in Human Primary Hypertension and Congestive Heart Failure	28	(9)
The Technique, Function and Structure	28	(1)
Remodelling	29	(2)
Endothelial Dysfunction	31	(2)
Forearm Veins in Primary Hypertension	33	(1)
Chronic Heart Failure	34	(3)
Pharmacology of Vascular Conduits for Coronary-Bypass Graft Surgery	37	(4)
Introduction	37	(1)
Internal Mammary Artery	38	(3)
Saphenous Vein	41	(1)
Human Vascular-to-Cardiac Tissue Selectivity of L-and T-Type VOCC Antagonists	41	(10)
References	47	(4)
Problems in Assigning Mechanisms: Reconciling the Molecular and Functional Pathways in α-Adrenoceptor-Mediated Vasoconstriction	51	(22)
M.J. Lew
Introduction	51	(1)
Coupling Mechanisms at the Molecular and Cellular Levels	52	(4)
α1-Adrenoceptors	52	(2)
α2-Adrenoceptors	54	(2)
Coupling Mechanisms in the Intact Animal	56	(3)
Reconciliation	59	(2)
Myogenic Activation	61	(1)
Autocrine and Paracrine Activation	62	(2)
Functional Antagonism	64	(2)
Adenylate Cyclase and α2-Adrenoceptors	66	(2)
Conclusions	68	(5)
References	69	(4)
Gs Protein-Coupled Receptors in Human Heart	73	(46)
A.J. Kaumann
Introduction	73	(1)
Receptor Subtypes	73	(1)
β-Adrenoceptor Subtypes	74	(6)
Comparison of β1- and β2-Adrenoceptors	74	(1)
Localisation	74	(1)
Function of β1- and β2-Adrenoceptors	74	(1)
Selective Coupling of β2-Adrenoceptors	75	(5)
Is There a Functional role for Cardiac β3-Adrenoceptors?	80	(2)
Evidence Against Cardiostimulation	80	(1)
Evidence for Cardiostimulation	81	(1)
Evidence for Cardiodepression	81	(1)
Evidence Against Cardiodepression	82	(1)
Cardiostimulant Effect Through the Putative β4-Adrenoceptor	82	(8)
Non-Conventional Partial Agonists	82	(2)
The Putative β4-Adrenoceptor Resembles - But Is Distinct from - the β3-Adrenoceptor	84	(3)
Which Endogenous Agonist for the Putative β4-Adrenoceptor?	87	(1)
The Putative β4-Adrenoceptor is a Special State of the β1-Adrenoceptor	88	(2)
5-HT4 Receptors	90	(5)
Coupling to a cAMP Pathway	90	(1)
5-HT4-like Receptors	91	(4)
Cross-Talk Between Cardiac Gs-Coupled Receptors, as Revealed by Chronic Blockade of β1-Adrenoceptors	95	(6)
Physiological, Pathophysiological and Therapeutic Relevance	101	(6)
β1-and β2-Adrenoceptors	101	(4)
Putative β4-Adrenoceptors	105	(1)
5-HT4 Receptors	105	(2)
Epilogue	107	(12)
References	108	(11)
Section II: New Theoretical Concepts and Molecular Mechanisms of Receptor Function
Kinetic Modeling Approaches to Understanding Ligand Efficacy	119	(28)
J.J. Linderman
Introduction	119	(1)
Background	120	(6)
Efficacy	120	(1)
Modeling	121	(1)
Equilibrium, Steady State, and Kinetic Models	122	(1)
Diffusion-Versus Reaction-Controlled Events	123	(1)
Model Structures and Dose-Response Curves	124	(2)
Parameters Contributing to Ligand Efficacy	126	(17)
The Lifetime of the Individual Receptor-Ligand Complex (1/kr)	126	(5)
The Receptor Desensitization Rate Constant kx	131	(3)
The Ligand Binding and Dissociation Rate Constants at Endosomal pH	134	(5)
Rate Constants in a Ternary Complex Model	139	(4)
Concluding Remarks	143	(4)
References	144	(3)
The Evolution of Drug-Receptor Models: The Cubic Ternary-Complex Model for G Protein-Coupled Receptors	147	(20)
T. Kenakin
P. Morgan
M. Lutz
J. Weiss
Receptor Models	147	(1)
The Ternary Complex Model of Receptor Function	148	(1)
Two-State Theory	149	(2)
The Extended Ternary Complex Model	151	(1)
The CTC Model	152	(3)
General Application of the Cubic Model	155	(8)
Evidence for the AriG Complex	158	(5)
Conclusion	163	(4)
References	163	(4)
Inverse Agonism	167	(16)
R. Bond
G. Milligan
M. Bouvier
Background	167	(1)
Overexpression, Spontaneous Activity and Inverse Agonism	168	(5)
Mutations and Diseases of Spontaneous Receptor Activity	173	(1)
Modulation of Receptor Function by Agonists and Inverse Agonists	174	(3)
Models	177	(1)
Summary and Conclusions	178	(5)
References	178	(5)
Efficacy: Molecular Mechanisms and Operational Methods of Measurement. A New Algorithm for the Prediction of Side Effects	183	(34)
T. Kenakin
Introduction	183	(1)
The Molecular Nature of Efficacy	183	(3)
Positive and Negative Efficacy	186	(2)
The Operational Measurement of Relative Efficacy	188	(13)
Binding Studies	188	(1)
Guanosine Triphosphate γS Shift	188	(3)
High-Affinity Selection Binding	191	(1)
Function	192	(3)
The Method of Furchgott	195	(1)
The Method of Stephenson	196	(1)
Comparison of Relative Maximal Responses	197	(4)
Limitations of Agonist Potency Ratios	201	(3)
Why Measure the Relative Efficacies of Agonists?	204	(2)
A Simple Algorithm for the Prediction of Agonist Side Effects Using Efficacy and Affinity Estimates	206	(9)
Therapeutic Versus Secondary Agonism: Side Effect Versus Coupling Constant Profiles	208	(2)
Algorithm for Calculation of Relative β50	210	(2)
Application of the Algorithm to β3-Adrenoceptor Agonists	212	(2)
Limitations of the Algorithm	214	(1)
Conclusions	215	(2)
References	215	(2)
A Look at Receptor Efficacy. From the Signalling Network of the Cell to the Intramolecular Motion of the Receptor	217	(44)
H.O. Onaran
A. Scheer
S. Cotecchia
T. Costa
Introduction	217	(1)
Biological Receptors and the Dualism of Affinity and Efficacy	217	(4)
Signal Transfer and Conformational Change in Membrane Receptor	217	(1)
The Distinction Between Affinity and Efficacy	218	(1)
Generality of the Concept of Efficacy in Functional Proteins	219	(1)
Asking the Questions	219	(1)
Two Flavours in the Definition of Efficacy: Biological and Molecular	220	(1)
Biological Definitions of Efficacy	221	(6)
The Nature of Signal Strength	221	(1)
Stimulus-Response Relationships	222	(1)
The Scale of Agonism and Antagonism	223	(2)
Steps of Signal Transduction and the Indeterminacy of Stimulus-Response Relationships	225	(2)
Molecular Definitions of Efficacy	227	(11)
A Molecular Link Between Affinity and Efficacy	227	(1)
Allosteric Equilibrium, Free-Energy Coupling, and Thermodynamic Definitions of Efficacy	227	(4)
Linkage Between Macroscopic Perturbations in the Receptor	231	(2)
Functional and Physical States in Proteins	233	(2)
Microscopic Interpretation of Allosteric Equilibrium	235	(3)
A Stochastic Model of Molecular Efficacy	238	(23)
Protein Motion and Fluctuations in Its Conformational Space	238	(1)
Probability Distribution of Microscopic States and Derivation of Macroscopic Constants	238	(3)
Probabilistic Interpretation of Ligand Efficacy	241	(3)
Relationship Between Physical States and Biological Function	244	(1)
Relationship Between Efficacy and Fluorescence Changes in β2-Adrenoceptors	244	(4)
Correlated Macroscopic Changes in Constitutively Active Adrenoceptors	248	(6)
References	254	(7)
Mechanisms of Non-Competitive Antagonism and Co-Agonism	261	(22)
D.G. Trist
M. Corsi
Non-Competitive Antagonism	261	(6)
Definition	261	(1)
Analysis of the Effect of Non-Competitive Antagonists	261	(6)
Co-Agonism	267	(12)
Theory as Applied to the Glutamate NMDA Receptor	267	(1)
Evidence for Co-Agonism	267	(1)
The Theory of Co-Agonism	268	(3)
The Effect of a Competitive Antagonist on the Response of Co-Agonism	271	(2)
The Effect of a Non-Competitive Antagonist on the Response of Co-Agonists	273	(1)
Experimental Data Supporting the Concept of Co-Agonism and Its Antagonism	274	(1)
Recombinant Experiments	274	(2)
Tissue Experiments	276	(2)
Implications of Co-Agonism	278	(1)
General Observations	279	(4)
References	280	(3)
Mechanisms of Receptor Activation and the Relationship to Receptor Structure	283	(30)
D.M. Perez
S.S. Karnik
Introduction	283	(1)
Common GPCR Structure/Function	283	(3)
Rhodopsin and bR Activation: Light as the Ligand	286	(5)
Direct Structural Information	287	(1)
The Activation Mechanism	288	(1)
How Retinal Binds	288	(1)
Salt-Bridge Constraining Factor: Movement of TM3 and TM6	289	(2)
AR Activation: Small Organics as Ligands	291	(7)
Important Binding Contacts of the Endogenous Ligands	291	(2)
Insights on How Epinephrine Activates	293	(1)
Release of Constraining Factors	293	(1)
Evidence for a Salt-Bridge as a Constraint: Movement of TM3 and TM7	294	(2)
Evidence for Multiple Activation States or Mechanisms	296	(1)
Evidence for Additional Constraining Factors: Movement of TM5 and TM6	296	(2)
Angiotensin-Receptor Activation: Peptides as Ligands	298	(4)
Peptide-Hormone GPCRs: How the Peptide Binds	298	(1)
The Ang-II Receptor	298	(1)
How AngII Peptides and Non-Peptides Bind	299	(1)
Insights into AngII Activation	300	(1)
Role of His256 in TM6	300	(1)
Release of Constraining Factors: Role of TM3	300	(2)
The Ties that Bind: Concluding Remarks	302	(11)
Conservation of Critical Binding Contacts and Resulting Helical Movements	302	(1)
Conservation of Switches that Control Attainment of the Active State(s)	302	(2)
References	304	(9)
Section III: New Technologies for the Study of Drug Receptor Interaction
The Assembly of Recombinant Signalling Systems and Their Use in Investigating Signaling Dynamics	313	(22)
S.M. Lanier
Introduction	313	(1)
Assembly of Recombinant Signaling Systems	314	(4)
Stable Transfection	314	(2)
Transient Expression Systems	316	(2)
Drug-Receptor Interactions in Recombinant Signaling Systems	318	(10)
Cell-Type-Specific Signaling Events	318	(5)
Influence of Accessory Proteins	323	(5)
Perspective	328	(7)
References	328	(7)
Insect Cell Systems to Study the Communication of Mammalian Receptors and G Proteins	335	(28)
R.T. Windh
A.J. Barr
D.R. Manning
Introduction	335	(3)
Selectivity of Receptor-G Protein Interactions	335	(1)
Effector Modulation as a Measure of G Protein Activity	336	(1)
Direct Measures of G Protein Activity	337	(1)
Insect Cell Expression Systems	338	(7)
Expression of Receptors in Insect Cell Lines	338	(4)
Interaction of Receptors with Endogenous G Proteins and Effectors	342	(1)
Quantitation of Coupling Using Radioligand Binding	343	(2)
Reconstitution of Mammalian Receptors and G Proteins: Reconstituted Properties of Ligand Binding	345	(5)
Influence of Heterotrimeric G Proteins on Binding of Agonists	346	(2)
Influence of Individual G Protein Subunits on Binding of Agonists	348	(1)
Characterization of Inverse Agonism	349	(1)
Reconstitution of Mammalian Receptors and G Proteins: G Protein Activation	350	(6)
Activation Following Co-Expression of Receptor and G Protein	350	(4)
Activation Following Addition of Purified G Protein to Membranes	354	(1)
Limitations and Technical Considerations	355	(1)
Conclusions	356	(1)
References	356	(7)
Altering the Relative Stoichiometry of Receptors, G Proteins and Effectors: Effects on Agonist Function	363	(28)
G. Milligan
Introduction	363	(4)
Background	363	(1)
Systems to Modulate GPCR--G Protein-Effector Stoichiometries	364	(1)
Cellular Distribution of Elements of G Protein-Coupled Signaling Cascades	365	(2)
GPCR-G Protein Fusion Proteins. A Novel Means to Restrict and Define the Stoichiometry of Expression of a GPCR and a G Protein α Subunit	367	(3)
G Protein-Coupled Receptors	370	(6)
Gsα	376	(4)
Effector Enzymes: Adenylyl Cyclase	380	(3)
Conclusions	383	(8)
References	383	(8)
The Study of Drug-Receptor Interaction Using Reporter Gene Systems in Mammalian Cells	391	(24)
D.M. Ignar
S. Rees
Reporter Systems	391	(10)
What is a Reporter Gene System?	391	(1)
Detection Methods	391	(1)
Enzymatic	391	(2)
Non-Enzymatic	393	(1)
Measurement of Intracellular Signaling	393	(1)
Inducible Reporter Genes	393	(1)
Responsive Promoters	393	(2)
Chimeric Transcription Factors	395	(2)
Reporter Proteins	397	(3)
Transient Versus Stable Expression of Reporter Genes	400	(1)
Transient Expression	400	(1)
Stable Expression	400	(1)
Use of Reporter Gene Systems in Pharmacology	401	(14)
Drug Discovery	401	(1)
Receptor Pharmacology	401	(1)
Use of Reporter Gene Assays to Assess Receptor Agonism	402	(1)
6CRE-Luciferase as a Reporter for Gαs-Coupled Receptor Signaling	402	(1)
Aequorin as a Reporter for Gαaq11-Coupled Receptor Signaling	402	(3)
The Use of a Ga14/Elk-1 Chimera to Report Opioid-Receptor-Like Receptor-1 Activation of MAPK	405	(1)
Use of Reporter Gene Assays to Evaluate Receptor Antagonism	405	(1)
Simultaneous Detection of Multiple Signals	405	(1)
Dual Reporter Assays	405	(2)
Combination of Reporter Assays with Other Assay Types	407	(2)
Measurement of constitutive Activity	409	(2)
Measurement of Efficacy	411	(1)
Assessment of New Signaling Pathways	411	(1)
References	412	(3)
Melanophore Recombinant Receptor Systems	415	(26)
C.K. Jayawickreme
M.R. Lerner
Introduction	415	(2)
Cellular Signaling in Melanophores	417	(6)
Signaling Pathways	417	(2)
Endogenous Receptor Signaling	419	(2)
Melatonin 1c Receptor	421	(1)
α-Melanocyte Stimulating Hormone (MSH) Receptor	421	(1)
Endothelin-C Receptor	422	(1)
Serotonin Receptor	422	(1)
β-Adrenoreceptor	422	(1)
VIP Receptor	423	(1)
Melanophore Assay Technology	423	(6)
Cell Culture and Related Techniques	423	(1)
Preparation of Cultures of Melanophore	423	(1)
Continuous Culturing of Xenopus laevis Melanophores	424	(1)
Receptor Expression	424	(1)
Preparation of Stable Melanophore Lines Expressing Exogenous Receptors	425	(1)
Signal Detection	425	(1)
Transmittance Reading	425	(1)
Digital Imaging	426	(1)
Screening Formats	427	(1)
Microtiter Well Format	427	(1)
Lawn Format	428	(1)
Receptor Cloning and Mutagenesis Studies	428	(1)
Receptor Studies and Applications	429	(6)
Characterization of Novel GPCRs	431	(1)
Lawn Format Screen System	432	(1)
Single Transmembrane Receptors	433	(1)
Receptor-Ligand Interaction Studies	434	(1)
Summary	435	(6)
References	435	(6)
Subject Index	441