The purpose of the present volume, the first of two on the pharmacology, biochemistry, and physiology of cyclic nucleotides, is to provide a comprehensive and up-to-date anthology on the nature and role of these important chemical regulators. Each of the chapters is the work of internationally known researchers who present a lucid and detailed review of their subject and not merely a single laboratory's viewpoint. The chapters emphasize critical assessments of the field rather than mere listings of experimental findings. By so doing, the contributors present the role of cyclic nucleotides in relationship to other intracellular regulators. Each chapter begins with a detailed summary to allow the reader to obtain a rapid overview of subsequent material. In addition, there are extensive bibliographies and a detailed subject index. Wherever pertinent, the chapters contain sections on drug mechanisms, physiological relevance, and disease processes. The Volume is divided into two sections, each beginning with an overview written by Professors T. W. RALL and P. GREENGARD, respectively. The first section focuses on the detailed pharmacology and chemistry of cyclic nucleotides, including their formation, degradation, measurement, and interaction with various modulatory agents, such as receptors and calcium. The second section is concerned with the biochemistry of protein phosphorylation, a process which appears to be one of the most important mechanisms for the intracellular expression of cyclic nucleotide action in eukaryotic cells.

Table of Contents:
Section I: Biochemistry of Cyclic Nucleotides.- 1 Formation and Degradation of Cyclic Nucleotides: An Overview.- A. Introduction.- B. Historical Review.- I. Discovery of Cyclic AMP.- II. Early Characterization of Adenylate Cyclase and Discovery of Cyclic Nucleotide Phosphodiesterase.- III. Discovery of Cyclic GMP and Guanylate Cyclase.- C. Regulation of Cyclic AMP Accumulation in Tissues.- I. Introduction.- II. Regulation of Cyclic AMP Formation.- 1. Basal and Hormone-Induced Accumilation of Cyclic AMP in Intact Cells.- 2. Effects of Guanyl Nucleotides and Cholera Toxin.- III. Regulation of Cyclic AMP Degradation.- IV. Potentiative Interactions and the Potential Role of Calcium.- D. Concluding Remarks.- References.- 2 Chemistry of Cyclic Nucleotides and Cyclic Nucleotide Analogs..- Overview.- A. Introduction to the Chemistry of Cyclic Nucleotides.- B. Synthesis of Cyclic Nucleotides and Cyclic Nucleotide Analogs.- I. Cyclic Nucleotides Related to Adenosine 3',5'-Cyclic Phosphate.- 1. Substituent Modification of the Purine Base.- 2. Structural Modification of the Carbohydrate Moiety.- 3. Structural Modification of the Cyclic Phosphate Moiety.- II. Cyclic Nucleotide Analogs Related to cAMP.- 1. 1-Deazapurine Cyclic Phosphates.- 2. 3-Deazapurine Cyclic Phosphates.- 3. 7-Deazapurine Cyclic Phosphates.- 4. 2-Azapurine Cyclic Phosphates.- 5. 8-Azapurine Cyclic Phosphates.- 6. Formycin Cyclic Phosphates.- 7. 1,N6-Ethenoadenosine Cyclic Phosphates.- III. Cyclic Nucleotides Related to Guanosine Cyclic Phosphate.- IV. Pyrimidine Cyclic Nucleotides.- V. Miscellaneous Cyclic Nucleotides.- C. Chemical and Physical Properties of Cyclic Nucleotides.- I. Hydrolytic Studies.- II. Spectral Measurements.- III. Ultraviolet Spectrophotometric Measurements.- D. Structure and Enzymic Activity Relationships of Certain Nucleoside 3?,5?-Cyclic Phosphates.- I. Structural Requirements for the Stimulation of Various Protein Kinases.- II. Structural Requirements for Stability Against Cyclic Nucleotide Phosphodiesterases.- III. Cyclic Nucleotides and Inhibition of Cellular Proliferation.- IV. Cyclic Nucleotides and Antiviral Effects.- V. Cyclic Nucleotide Analogs and Cardiovascular Effects.- E. List of Abbreviations Used.- References.- 3 Coupling of Receptors to Adenylate Cyclases.- Overview.- A. Introduction.- B. Guanine Nucleotide Regulation of Adenylate Cyclases.- C. Regulation of Coupling of Hormone Receptor to Adenylate Cyclase.- D. Receptor Regulation and Receptor Forms.- I. Effects of Guanine Nucleotides on Binding of Hormone to Receptor.- II. Definition of an Active Form of Receptor.- III. Number of Sites Involved in Coupling.- E. Concluding Remarks and Future Outlooks.- I. Roles of Mg++.- II. Inhibitory Effects of Hormones.- III. Is GTPase Indeed an Integral Part of Adenylate Cyclases?.- IV. Are Levels of N Regulated?.- V. Molecular Basis for Coupling: Size Studies and Subunit Interactions.- References.- 4 Acute and Chronic Modulation of the Responsiveness of Receptor-Associated Adenylate Cyclases.- Overview.- A. Introduction.- I. Historical Perspective.- II. Scope of the Review.- III. The Structure and Function of Adenylate Cyclase.- B. Agonist-Induced Decreases in the Response of Cells to Hormones.- I. Early Observations.- II. Agonist-Induced Desensitization of Intact Cells.- 1. General Characteristics of Agonist-Induced Desensitization.- 2. Effects of Cyclic AMP and Cyclic AMP Analogs.- 3. Whole Cell Analysis of Rates of Cyclic AMP Synthesis and Degradation.- 4. Conclusions Based on Studies with Whole Cells.- III. Agonist-Induced Changes in Adenylate Cyclase and Hormone Binding.- IV. Separation of Native and Desensitized Beta-Adrenergic Receptors..- V. Catecholamine-Induced Desensitization in C6-2B Glioma Cells.- VI. Agonist-Induced Desensitization in Cell-Free Preparations.- VII. Desensitization of Gonadotropin Receptor-Linked Adenylate Cyclase.- C. Receptor-Mediated Inhibition of Adenylate Cyclase Activity.- D. Physiological Significance of the Regulation of Responsiveness to Hormones.- References.- 5 Guanylate Cyclase: Regulation of Cyclic GMP Metabolism..- Overview.- A. Introduction.- B. Guanylate Cyclase.- I. Activities in Various Tissues and Subcellular Distribution.- II. Altered Enzyme Activity and Subcellular Distribution.- III. Kinetic and Physical Properties of the Multiple Forms.- 1. Molecular Size and Subunits.- 2. Effect of GTP and Other Nucleotides.- 3. Effects of Cations.- 4. Antibodies Against Guanylate Cyclase.- IV. Activation.- 1. Detergents.- 2. Spontaneous Activation.- 3. Effects of Azide and Nitro Compounds.- 4. Oxidation and Hydroxyl Radical.- 5. Fatty Acids and Their Metabolites.- 6. Phospholipase and Lipoxygenase.- 7. Effects of Proteins.- V. Altered Kinetic Properties with Activation.- 1. Cation Dependence.- 2. Effects of Nucleotides.- 3. Formation of Cyclic AMP.- VI. Inactivation.- 1. Hemeproteins.- 2. Other Inhibitors.- C. Cyclic GMP Levels in Tissues.- I. Effects of Hormones and Autacoids.- 1. Effects of Pituitary Hormones and Thyroid Function.- 2. Choline Esters.- 3. Histamine.- 4. Alpha-Adrenergic Agonists.- 5. Insulin.- 6. Ionophore A-23187.- 7. Other Agents.- II. Effects of Nitro Compounds and Nitric Oxide.- III. Effect of E. Coli Heat-Stable Enterotoxin.- D. Conclusion and Hypotheses.- References.- 6 Cyclic Nucleotide Phosphodiesterase..- Overview.- A. The Phosphodiesterase Reaction and Assay.- B. Multiple Forms of Phosphodiesterase.- I. Identification and Separation.- II. Properties.- III. In vitro Alterations and Possible Interconversions.- C. Localization.- D. Activators and Inhibitors.- I. Cyclic Nucleotides as Effectors of Enzyme Activity.- II. Other Effectors.- E. Physiological Roles of the Cyclic Nucleotide Phosphodiesterases.- I. Development.- II. Transformation and Malignancy.- III. Insulin and Other Metabolic Hormone Actions.- IV. Neurotransmission.- F. Prognosis.- References.- 7 Calmodulin Regulation of Cyclic AMP Metabolism..- Overview.- A. Introduction.- B. Calmodulin.- I. Properties of Calmodulin.- II. Preparation and Assay of Calmodulin.- III. Calmodulin-Regulated Cellular Processes.- IV. Mechanism of Action.- C. Adenylate Cyclase.- I. Ca2+ Sensitivity of Adenylate Cyclase.- II. Regulation of Adenylate Cyclase by Calmodulin.- D. Cyclic Nucleotide Phosphodiesterase.- I. Multiple Forms of Phosphodiesterase.- II. Ca2+-Dependent Phosphodiesterase.- E. Biological Significance.- References.- 8 Radioimmunoassay Techniques for Cyclic Nucleotides.- Overview.- A. Introduction.- B. Preparation and Characterization of Antisera.- C. Preparation and Purification of [125I]-Labeled Cyclic Nucleotides.- D. Preparation of Samples for Assay.- E. Acetylation or Succinylation.- F. Assay Procedure.- G. Data Analysis.- H. Evaluation of Assay Validity.- I. Total Automation of the Cyclic AMP and Cyclic GMP Assay with the Gamma-Flo Assay System.- References.- 9 Immunocytochemistry of Cyclic Nucleotides and Their Kinases..- Overview.- A. Introduction.- B. Principles.- C. Techniques.- D. Cyclic Nucleotide Immunocytochemistry.- I. Preparation of Antisera.- II. Testing of Antisera for Specificity of Staining.- III. Methodological Considerations.- E. Protein Kinase Immunocytochemistry.- I. Purification of Immunogens.- II. Preparation and Selection of Antisera.- III. Characterization of Antisera for Immunocytochemistry.- 1. Cyclic AMP-Dependent Protein Kinases and Their Subunit Antisera.- 2. Cyclic GMP-Dependent Protein Kinase Antisera.- IV. Determination of Specificity of Protein Kinase Antisera by Radio-immunoassay.- V. Testing of Antisera for Specificity of Staining.- VI. Methodological Considerations.- 1. Crossreactivity of Antibodies for Holoenzyme and Their Subunits.- 2. Avidity of an Antiserum for Available Determinants in Tissue.- 3. Losses and Redistribution of Protein Kinases.- 4. Species Specificity of Protein Kinase Immunocytochemistry.- F. Future Perspectives.- References.- Section II: Biochemistry of Protein Phosphorylation.- 10 Protein Phosphorylation: An Overview.- References.- 11 Cyclic AMP-Dependent Protein Phosphorylation.- Overview.- A. Introduction.- B. History of Discovery of cAMP-Dependent Protein Kinase.- C. Nomenclature and General Activation Mechanism of Protein Kinase.- D. Isolation of cAMP-Dependent Protein Kinase.- E. Physical Properties of Protein Kinase.- F. Naturally Occurring Mechanisms Modulating Protein Kinase Activity.- I. Heat-Stable Protein Inhibitors.- 1. Muscle Inhibitor.- 2. Testis Inhibitor.- II. Nucleotide Effects.- 1. cAMP.- 2. ATP.- III. ATP-Dependent Phosphorylations.- 1. Autophosphorylation of RII.- 2. Phosphorylation of RI.- G. Substrates of cAMP-Dependent Protein Kinase.- I. Primary Sequence Determinants.- II. Higher Order of Structure Effects.- H. Physiological Effects of Phosphorylation.- I. Criteria for Evaluating Physiological Roles of Cyclic Nucleotide-Dependent Phosphorylation Reactions.- II. Methods of Demonstrating Criteria, Problems Encountered, and Interpretation of Results.- III. Effects of Phosphorylation on the Activity or Function of the Substrate.- IV. Proteins of Known Function as Substrates.- I. Current Areas of Research.- I. Current Research on cAMP-Dependent Protein Kinase.- 1. Stoichiometry and Properties of cAMP Binding.- 2. Mechanism of Activation and Inactivation.- 3. Compartmentalization.- 4. Genetic Studies, Mutant Cell Lines.- 5. Differential Regulation of R and C.- II. Current Research on Substrates of Protein Kinase.- 1. Multiple Phosphorylations.- 2. Recently Identified Substrates.- References.- 12 Cyclic GMP-Dependent Protein Phosphorylation.- Overview.- A. Introduction.- B. Enzymology.- I. Distribution.- II. Molecular Properties and Mechanism of Action of Cyclic GMP.- III. General Catalytic Properties.- C. Substrate Proteins.- I. Autophosphorylation of G-PK.- II. Histone.- III. Non-Histone Proteins.- 1. Phosphorylase Kinase.- 2. Glycogen Synthase.- 3. Hormone-Sensitive Lipase and Cholesterol Ester Hydrolase.- 4. Pyruvate Kinase.- 5. Troponin Inhibitory Subunit (TN-I).- 6. Other Proteins.- IV. Membrane and Cytoplasmic Proteins.- D. Factors Influencing Protein Phosphorylation.- I. Stimulatory Modulator.- II. Polyanions, Polycations, and Basic Polypeptides.- E. Physio-Pathologic Aspects of the G-PK System.- F. Concluding Remarks.- References.- 13 Calcium-Dependent Protein Phosphorylation.- Overview.- A. Introduction.- B. Role of Calcium in Regulation of Carbohydrate Metabolism.- C. Role of Calcium in Regulation of Protein Phosphorylation in Intact Systems.- I. Calcium-Dependent Protein Phosphorylation in Brain.- II. Calcium-Dependent Protein Phosphorylation in Mast Cells..- III. Regulation of Protein Phosphorylation in Isolated Hepatocytes.- D. Calmodulin- and Calcium-Dependent Protein Kinase in Biological Membranes.- I. Calcium-Dependent Protein Phosphorylation in Isolated Brain Membranes.- II. Identification of Protein Kinase Activator as Calmodulin.- III. Characteristics of Protein Kinase Activity.- IV. Function of Calcium-Dependent Protein Phosphorylation in Brain.- V. Regulation of Protein I Phosphorylation by cAMP and by Calcium.- E. Widespread Occurrence of Calmodulin- and Calcium-Dependent Protein Phosphorylation in Biological Membranes.- F. Myosin Light Chain Kinase in Skeletal Muscle, Smooth Muscle, and Non-Muscle Cells.- I. Myosin Light Chain Kinase in Skeletal Muscle.- II. Myosin Light Chain Kinase in Smooth Muscle.- III. Myosin Light Chain Kinase in Non-Muscle Cells.- IV. Mechanism of Activation of Myosin Light Chain Kinase.- V. Physiological Function of P-Light Chain Phosphorylation.- G. Phosphorylase Kinase.- I. Regulation of Phosphorylase Kinase by Calmodulin.- II. Activation of Phosphorylase Kinase by Troponin C.- III. Regulation of Phosphorylase Kinase in Vitro.- IV. Regulation of Phosphorylase Kinase in Vivo.- H. Phospholipid-and Calcium-Dependent Protein Kinase.- I. Irreversible Activation of Protein Kinase C by Calcium-Dependent Protease.- II. Reversible Activation of Protein Kinase C by Calcium and Membrane Factor.- I. Concluding Comments.- References.- 14 Photoaffinity Labeling of Cyclic AMP-Dependent and Cyclic GMP-Dependent Protein Kinases.- Overview.- A. Introduction.- I. Principles of Affinity Labeling and Photoaffinity Labeling.- II. Brief Review of cAMP- and cGMP-Receptor Proteins.- B. Photoaffinity Analogues of cAMP and cGMP.- I. Available Photoaffinity Analogues of cAMP and cGMP.- II. Synthesis and Characterization of 8-N3-[32P] cAMP.- III. Procedures for Photoaffinity Labeling and Other Methods to Assay cAMP- and cGMP-Dependent Protein Kinases.- C. Studies with Purified cAMP-Dependent Protein Kinase.- I. Photoaffinity Labeling of Regulatory Subunit.- II. Photoaffinity and Affinity Labeling of Catalytic Subunit.- D. Application of Photoaffinity Labeling to Crude Systems.- E. Specific Examples of the Usefulness of the Photoaffinity Label 8-N3-[32P] cAMP.- I. Distribution and Differences of Type I and Type II cAMP-Dependent Protein Kinase.- II. Proteolytic Fragments of cAMP-Dependent Protein Kinases.- III. Subcellular Distribution of cAMP-Dependent Protein Kinases and cAMP-Receptor Proteins.- IV. Regulation of cAMP-Dependent Protein Kinase and cAMP-Receptor Proteins.- F. Photoaffinity Labeling of cGMP-Dependent Protein Kinase.- G. Problem Areas.- H. Conclusions.- References.- 15 Nuclear Protein Phosphorylation and the Regulation of Gene Expression.- Overview.- A. Introduction.- B. Nuclear Cyclic Nucleotide-Dependent Protein Kinase Activities.- I. Cyclic AMP-Dependent Protein Kinase.- 1. Translocation to Nuclei.- 2. Endogenous Nuclear Cyclic AMP-Dependent Protein Kinase.- II. Cyclic GMP-Dependent Protein Kinase.- III. Cytochemical Localizations of Protein Kinase Activities.- C. Histone Phosphorylation.- I. H1 Phosphorylation.- 1. Cyclic Nucleotide-Dependent H1 Phosphorylation.- 2. Cyclic Nucleotide-Independent H1 Phosphorylation.- II. Phosphorylation of Histones Other than H1.- III. Role of H1 Phosphorylation in Altering Nucleosome Structure.- D. Non-Histone Protein Phosphorylation.- I. Phosphorylation of Eukaryotic DNA-Dependent RNA Polymerases.- II. Phosphorylation of HMG Proteins.- E. Phosphorylation of Ribosomal Proteins.- F. Protein Phosphorylation and Viral Transformation.- G. Functional Aspects of Protein Kinase Substrates: A Procedure for Isolating Newly-Phosphorylated Polypeptides.- H. Protein Phosphorylation as a Means of Altering the Genetic Program: The Problem of Specificity.- I. Conclusions.- References.