Fundamentals of Intracellular Calcium

Anthony K. Campbell, PhD, FLS, FLSW is Professor in the School of Pharmacy and Pharmaceutical Sciences, Cardiff University. He has studied intracellular calcium for over 40 years, and is a world authority on bioluminescence, developing the use of genetically engineered bioluminescence to measure chemical processes in live cells, particularly Ca2+. One of his inventions, using chemiluminescence, is now used in several hundred million clinical tests per year worldwide. He has published 9 books, and over 250 internationally peer-reviewed papers on intracellular calcium, bioluminescence, lactose and food intolerance, and Darwin.

• About the Author xviiPreface xixList of Acronyms xxiiiAbout the Companion Web Site xxxv1 Calcium is Special 11.1 Calcium and Everyday Events 11.2 Discovery of Calcium 21.3 A Natural History of Calcium 31.3.1 Calcium by the Sea 31.3.2 Calcium in Your Wake-Up Call 41.4 The Elements of Life 51.5 Natural Occurrence of Calcium 71.5.1 Isotopes of Calcium 71.5.2 Geology of Calcium 71.5.3 Calcium Outside Cells 81.5.4 Calcium Inside Cells 101.6 Requirement of Cells for Ca2+ 121.6.1 Calcium in External Fluids 121.6.2 Requirement of Cell Types for Calcium 121.7 The Four Biological Roles of Calcium 131.7.1 Structural 131.7.2 Electrical across Biological Membranes 131.7.3 Extracellular Cofactor 141.7.4 Intracellular Regulator 141.8 The Puzzle about Ca2+ Inside Cells 171.9 How Important Intracellular Calcium has been in Science 181.10 Darwin and Intracellular Ca2+ 191.11 The Scene Set 211.12 ‘Ja Kalzium, das ist alles!’ 211.13 Calcium – The Fundamentals 22Recommended Reading 23Books 23Reviews 23Articles 232 Intracellular Calcium – Principles 252.1 Ca2+ and the Concept of Cell Signalling 252.1.1 Primary Signals and Intracellular Ca2+ 252.1.2 The Source of Intracellular Ca2+ 272.2 The Problem 292.3 Some Key Questions 292.3.1 Question 1 – What is the Primary Stimulus and Secondary Regulator(s)? 292.3.2 Question 2 – What is the Intracellular Signal? 302.3.3 Question 3 – How do the Secondary Regulators Work? 302.4 Types of Intracellular Ca2+ Signal 312.4.1 Category of Cell Based on the Ca2+ Signal 312.4.2 Types of Ca2+ Signal 322.4.3 Ca2+ Sparks, Puffs and Sparklets 332.5 Rubicon Principle 342.5.1 Digital Versus Analogue Cell Events 342.5.2 The Ca2+ Signaling Toolkit 362.5.3 Amplification Mechanisms 362.6 Key Experiments to Answer Key Questions 382.6.1 Raising Cytosolic Free Ca2+ Artificially 382.6.2 Measurement of Free Ca2+ in Live Cells 382.6.3 How the Ca2+ Signal is Generated 392.6.4 Identifying the Ca2+ Target 392.6.5 How Binding of Ca2+ to its Target Causes the Cell Event 392.6.6 How the Cell Returns to Rest 402.7 Nomenclature – How Things are Named 402.7.1 English and Etymology 402.7.2 Organisms 412.7.3 Acronyms, Cells and Biochemistry 412.7.4 Genes 422.7.5 Proteins and Protein Sequences 432.7.6 Enzymes 432.7.7 Ion Channels 442.7.8 Agonists and Antagonists 472.7.9 Chemicals 472.7.10 Toxins 482.7.11 Drugs 482.7.12 Ca2+ Indicators 492.7.13 Units 502.8 Model Systems 502.9 Darwin, Wallace and Intracellular Ca2+ 512.9.1 Natural Selection in Real Time 512.9.2 Small Change by Small Change 522.10 New Knowledge 542.11 Conclusions 55Recommended reading 55Books 55Reviews 55Papers 563 A Century plus of Intracellular Ca2+ 573.1 Background 573.1.1 Pathway of Discovery and Invention for Intracellular Ca2+ 573.1.2 Model Systems 583.2 Why Study the History of Science? 593.3 The Tale of Three Pioneers and What Followed 593.3.1 Experiments of Sydney Ringer (1836–1910) 593.3.2 The Vision of Lewis Victor Heilbrunn (1892–1959) 613.3.3 Setsuro Ebashi (1922–2006): Pioneer of Intracellular Ca2+ in Muscle Contraction 633.4 Ca2+ as an Intracellular Regulator 643.4.1 Cell Theory 653.4.2 Origin of the Use of Ca2+ 653.4.3 Manipulation of Ca2+ 663.4.4 Measurement and Location of Free Ca2+ in Live Cells 663.4.5 Identification of the Components Responsible for Regulating Free Ca2+ Inside Cells 713.4.6 Discovery of Plasma Membrane Ca2+ Pumps and Transporters 713.4.7 Discovery of How Ca2+ is Released from the Sarco-/Endo-Plasmic Reticulum 723.4.8 Discovery of IP3 and its Receptor 733.4.9 Discovery of the Ryanodine Receptor 733.4.10 Discovery of Store-Operated Calcium Entry (SOCE) 743.4.11 Discovery of Ca2+ Channels in the Plasma Membrane 763.4.12 Discovery of TRP Channels 783.4.13 Discovery of G-Proteins 783.4.14 Ca2+ Targets Inside Cells and How They Work 783.4.15 Intracellular Ca2+ in Plants 803.4.16 Intracellular Ca2+ in Bacteria 813.4.17 Pathology of Intracellular Ca2+ 813.5 Conceptual Development of Ca2+ as an Intracellular Regulator 823.6 Summary 84Recommended Reading 86Books 86Reviews 86Research Papers 864 How to Study Intracellular Ca2+ as Cell Regulator 894.1 Pathway to Discover the Role of Intracellular Ca2+ in a Cell Event 894.2 Manipulation of Intracellular Ca2+ 904.3 Measurement of Intracellular Free Ca2+ 924.3.1 Absorbing Dyes 934.3.2 Fluorescent Ca2+ Indicators 944.3.3 Calibration of Fluors 964.3.4 Ca2+-Activated Photoproteins 984.3.5 Calibration of Ca2+-Activated Photoproteins 1024.3.6 Ca2+ Indicators from Engineered GFP 1054.3.7 Other Free Ca2+ Indicators for Whole Organs and Organisms 1094.4 Detecting and Imaging Photons 1094.5 Measurement of Total Cell Ca2+ 1134.6 Calcium Buffers 1134.7 Measurement of Ca2+ Fluxes 1154.8 How to Study Ca2+ Channels 1164.9 Discovering How the Rise in Cytosolic Free Ca2+ Occurs and thenReturns to Rest 1174.10 Discovering the Intracellular Ca2+ Target and How it Works 1184.11 Other Ions 1184.12 Summary 119Recommended Reading 120Books 120Reviews 120Research Papers 1215 How Ca2+ is Regulated Inside Cells 1235.1 Principles 1235.2 How Resting Cells Maintain their Ca2+ Balance 1265.3 Electrophysiology of Intracellular Ca2+ 1275.4 Primary Stimuli Producing a Cytosolic Ca2+ Signal 1305.5 Plasma Membrane Ca2+ Channels 1315.5.1 Voltage-Gated Ca2+ Channels 1315.5.2 TRP Channels 1365.5.3 Receptor-Activated Ca2+ Channels 1375.5.4 Mechanosensitive Channels 1385.5.5 Store-Operated Calcium Entry (SOCE) 1385.5.6 Ca2+ Receptor 1395.6 Regulation of Intracellular Ca2+ by, and within, Organelles 1405.6.1 Endo- and Sarco-Plasmic Reticulum (ER/SR) 1405.6.2 Mitochondrial Ca2+ 1465.6.3 Lysosomal and Endosomal Ca2+ 1485.6.4 Secretory Vesicle Ca2+ 1495.6.5 Peroxisomal Ca2+ 1495.6.6 Control of Ca2+ by the Golgi 1505.6.7 Nucleus and Ca2+ 1505.6.8 Plant Organelles and Ca2+ 1515.6.9 Acidocalcisomes 1525.7 Second Messengers and Regulation of Ca2+ Signalling in the Cytosol 1525.8 Pore Formers and Intracellular Ca2+ 1545.9 Gap Junctions and Connexins 1545.10 Other Ion Channels and Ca2+ 1555.11 Summary – How Ca2+ is Regulated Inside Cells 156Recommended Reading 157Books 157Reviews 157Research Papers 1586 How Ca2+ Works Inside Cells 1616.1 Biological Chemistry of Ca2+ 1616.1.1 The Special Biological Chemistry of Ca2+ 1616.1.2 Key Chemical Properties of Ca2+ 1646.1.3 Ca2+ Ligands 1646.1.4 Solvation 1666.1.5 Activity Coefficients 1676.1.6 Fractional Ca2+ Binding After a Ca2+ Signal 1676.1.7 Kinetics 1696.1.8 Diffusion 1696.1.9 Solubility 1706.2 Ca2+-Binding Proteins 1706.2.1 Ca2+-Binding Proteins Inside Cells 1706.2.2 Proteins with the EF-Hand Motif 1716.2.3 Proteins with the C2 Motif 1776.2.4 Proteins with a Cluster of Acidic Residues 1786.2.5 Proteins Forming a Cluster of Oxygen Ligands 1796.3 Ca2+ and Other Intracellular Signals 1826.3.1 Cyclic Nucleotides and Ca2+ 1826.3.2 Ca2+ and Protein Kinase C (PKC) 1846.3.3 Nitric Oxide and Ca2+ 1846.3.4 Ca2+, Inositol Phosphates and Other Intracellular Signals 1856.4 Ca2+ and Monovalent Ions 1856.4.1 Intracellular Ca2+ and K+ Conductance 1866.4.2 Intracellular Ca2+ and Na+ Conductance 1876.4.3 Ca2+ and Chloride Channels (CaCC) 1886.5 Other Cations and Intracellular Ca2+ 1886.6 Anions and Intracellular Ca2+ 1896.7 Summary 189Recommended Reading 191Books 191Reviews 191Research Papers 1927 How Ca2+ Regulates Animal Cell Physiology 1937.1 Principles 1937.2 Ca2+ and How Nerves Work 1947.3 Muscle Contraction 1967.4 Chemotaxis and Ca2+ 2027.5 Intracellular Ca2+ and Secretion 2037.6 Ca2+ and Endocytosis 2097.7 Ca2+ and Phagocytosis 2107.8 Intracellular Ca2+ and Intermediary Metabolism 2107.9 Intracellular Ca2+ and Cell Growth 2127.9.1 Principles 2127.9.2 Cell Cycle and Ca2+ 2127.9.3 Fertilisation and Intracellular Ca2+ 2147.9.4 Differentiation and Intracellular Ca2+ 2167.10 Intracellular Ca2+ and the Immune Response 2167.11 Intracellular Ca2+ and Vision 2177.11.1 Ca2+ and Vertebrate Vision 2187.11.2 Ca2+ and Invertebrate Vision 2197.12 Intracellular Ca2+ and Other Senses 2197.13 Ca2+ and Bioluminescence 2207.14 Intracellular Ca2+ and Gene Expression 2227.15 Summary 223Recommended Reading 224Books 224Reviews 225Research Papers 2258 Intracellular Ca2+, Microbes and Viruses 2278.1 The Puzzle 2278.2 What Microbes Do 2288.3 Indirect Evidence of a Role for Intracellular Ca2+ in Bacteria 2288.4 Direct Evidence for a Role of Intracellular Ca2+ in Bacteria 2308.5 How Much Ca2+ is there in Bacteria? 2318.6 How Bacteria Regulate Their Intracellular Ca2 2328.6.1 Ca2+ Influx into Bacteria 2338.6.2 Ca2+ Efflux out of Bacteria 2368.7 Regulation of Bacterial Events by Intracellular Ca2+ 2368.7.1 Ca2+ and Growth of Bacteria 2368.7.2 Ca2+ and Bacterial Movement 2368.7.3 Quorum Sensing and Gene Expression 2408.7.4 Ca2+ and Bacterial Metabolism 2408.7.5 Bacterial Defence – Dormancy, Spore Formation and Germination 2418.7.6 Bacterial Infection – Virulence, Competence and Defence 2428.7.7 Development of Bacterial Structures 2438.7.8 Ca2+ and Gene Expression in Bacteria 2458.7.9 Uptake of Nucleic Acid by Bacteria 2458.7.10 Bacterial Metabolic Toxin Hypothesis 2468.7.11 Intracellular Ca2+ in Bacteria – Conclusions 2468.8 Role of Intracellular Ca2+ in Archaea 2478.9 Intracellular Ca2+ and Viruses 2498.9.1 Eukaryotic Viruses 2498.9.2 Bacterial Viruses – Bacteriophages 2518.10 Intracellular Ca2+ and Eukaryotic Microbes 2518.10.1 Yeast 2528.10.2 Paramecium and Related Ciliates 2548.10.3 Slime Moulds 2558.10.4 Luminous Radiolarians 2578.11 Summary 257Recommended Reading 258Books 258Reviews 259Research Papers 2599 Role of Intracellular Ca2+ in Plants and Fungi 2619.1 Role of Ca2+ in Plants 2619.2 What Stimulates Plants? 2629.2.1 Plant Cell Stimuli 2629.2.2 Plant Hormones 2639.2.3 Intracellular Signals in Plants 2659.3 Requirement of Plants for Ca2+ 2659.4 Where Ca2+ is Stored in Plants 2669.5 Measurement of Cytosolic Free Ca2+ in Plants 2679.6 Components of Ca2+ Signalling in Plants 2719.6.1 Ca2+ Pumps and Exchangers 2719.6.2 Ca2+ Channels in the Plasma Membrane 2719.6.3 Plant Organelles and Ca2+ 2729.6.4 Ca2+-Binding Proteins in Plants 2739.6.5 Ca2+-Sensitive Genes in Plants 2749.7 How Intracellular Ca2+ Provokes Cellular Events in Plants 2749.7.1 Light and Intracellular Ca2+ in Plants 2759.7.2 Control of Opening and Closing of Stoma/Stomata 2759.7.3 Wind and Mechanical Stimulation of Plants 2769.7.4 Gravity Sensing and Growth 2769.7.5 Fertilisation, Germination and Differentiation 2779.7.6 Legumes 2789.7.7 Intermediary Metabolism 2789.7.8 Transport by Phloem 2799.7.9 Defence Against Stress 2799.8 Fungal Elicitors 2809.9 Apoptosis 2819.10 Intracellular Ca2+ and Plant Pathology 2819.11 Ca2+ in Mosses, Liverworts and Ferns 2819.12 Ca2+ in Fungi 2829.12.1 Fungi and Intracellular Ca2+ 2829.12.2 Intracellular Ca2+ and Yeast 2839.12.3 Lichens 2849.13 Ca2+ and Slime Moulds 2849.14 Summary 284Recommended Reading 285Books 285Reviews 285Research Papers 28610 Pathology of Intracellular Ca2+ 28710.1 What is Pathology? 28710.2 Types of Pathology 28810.3 Intracellular Ca2+ – Friend or Foe? 28910.4 Intracellular Ca2+ and Cell Death 29010.4.1 Necrosis 29110.4.2 Apoptosis 29210.4.3 Autophagy 29410.4.4 Lysis 29510.4.5 Cell Death Conclusions 29510.5 Genetic Abnormalities in Ca2+ Signalling Proteins 29710.5.1 Ca2+ Channelopathies 29810.5.2 Ca2+ Pumpopathies 29810.5.3 Mutations in ER Ca2+ Release Proteins 29910.5.4 Mutations in Ca2+ Target Proteins 30010.5.5 Proteins Associated with Ca2+ Signalling 30010.6 Oxygen and Cell Pathology 30010.6.1 Ca2+ Paradox 30010.6.2 Oxidative Damage and Intracellular Ca2+ 30210.7 Inappropriate Ca2+ Signalling 30210.7.1 Cramp 30310.7.2 Immune System and Other Organs in Disease 30310.7.3 Bacterial Metabolic Toxins 30410.8 ER Stress Response 30610.9 Summary 308Recommended Reading 309Books 309Reviews 309Research Papers 31011 Pharmacology of Intracellular Ca2+ 31311.1 Background 31311.2 Pharmacological Targets for Intracellular Ca2+ 31611.3 Drugs Used Clinically on Intracellular Ca2+ 31711.4 Anaesthetics 31711.5 Ca2+ Channel Effectors 31911.5.1 Classes of Ca2+ Channel Blocker 31911.5.2 Dihydropyridines 32111.5.3 Phenylalkylamines 32211.5.4 Benzothiazapines 32211.6 Hypertension 32311.7 Arrhythmia, Tachycardia and Bradycardia 32311.8 Angina 32411.9 Heart Failure 32411.10 Agents that Affect Adrenergic Receptors 32511.11 Cardiac Glycosides 32611.12 Benzodiazapines 32611.13 Anti-Psychotic Drugs 32711.14 Stimulants and Drugs of Abuse 32711.15 Analgesics 32711.16 Anti-Depressants and Manic Depression 32911.17 Diabetes 32911.18 Muscle Relaxants 32911.19 Anti-Allergics and Anti-Immune Compounds 33011.20 Xanthines 33111.21 Substances Used Experimentally to Interfere with Intracellular Ca2+ 33111.21.1 Ca2+ Buffers and Ionophores 33211.21.2 Ca2+ Channels 33211.21.3 Agents which Open or Close Voltage-Gated Ca2+ Channels 33211.21.4 IP3 Receptor Activators and Blockers 33311.21.5 Ryanodine Receptor Agonists and Antagonists 33511.21.6 Plasma Membrane Ca2+ Pump and Exchanger Inhibitors 33511.21.7 SERCA Pump Inhibitors 33511.21.8 Compounds which Affect SOCE 33611.21.9 Mitochondrial Blockers 33611.21.10 Ca2+ Target Inhibitors 33711.22 Natural Toxins and Poisons 33911.22.1 Natural Stings, Bites and Other Toxin Events 33911.22.2 Cone Snail Toxins – Conotoxins 34011.22.3 Spider Toxins – Agatoxins 34211.22.4 Scorpion Toxins 34211.22.5 Pufferfish and Octopus Toxins 34311.22.6 Jellyfish Sting Toxins 34311.22.7 Hymenopteran Stings and Bites 34411.22.8 Snake Venoms 34511.23 Plant Toxins and Intracellular Ca2+ 34511.24 Drugs and the Ca2+ Receptor 34611.25 Bacteria 34711.26 Ions and Intracellular Ca2+ 34711.27 Antibodies and Intracellular Ca2+ 34811.28 Manipulation of mRNA 34911.29 Summary 349Recommended Reading 350Books 350Reviews 350Research Papers 35112 Darwin and 4000 Million Years of Intracellular Ca2+ 35312.1 Darwin and Calcium 35312.2 Evolution and Ca2+ 35712.3 What is Evolution? 35712.3.1 The Word Evolution 35712.3.2 The Process 35812.3.3 Sequence of Earth’s Evolution 35912.3.4 Ca2+ and Primeval Life 36012.3.5 Ca2+ and the Origin of the Three Fundamental Cell Types 36012.3.6 Timescale 36112.4 Evolution of Ca2+ Signalling 36212.4.1 Origin of Ca2+ Signalling 36212.4.2 Membrane Potential 36212.4.3 Evolution of Ca2+ Signalling Based on Protein Sequences 36412.4.4 Evolution of Ca2+-Binding Sites 36812.4.5 Origin of the EF-Hand 36912.5 Darwin and Knock-Outs 37112.6 Summary 373Recommended Reading 375Books 375Reviews 375Research Papers 37613 They Think It’s All Over 37913.1 Calcium and the Beauty of Nature 37913.2 What We Know About Intracellular Ca2+ 38013.2.1 Principles 38013.2.2 The Pathway of Discovery 38313.3 What We Don’t Know About Intracellular Ca2+ 38513.4 Intracellular Ca2+ at School and University 38713.5 The Inspiration of Intracellular Ca2+ 38813.6 Communicating the Story of Intracellular Ca2+ to Others 39113.7 The End of the Beginning 396Recommended Reading 398Books 398Reviews 398Research Papers 398Subject Index 399