{"product_id":"pumps-channels-and-transporters-9781118858806","title":"Pumps Channels and Transporters","description":"\u003cb\u003eBook Synopsis\u003c\/b\u003e\u003cbr\u003eDescribes experimental methods for investigating the function of pumps, channels and transporters\u003cbr\u003e \u003cul\u003e \u003cli\u003eCovers new emerging analytical methods used to study ion transport membrane proteins such as single-molecule spectroscopy\u003c\/li\u003e \u003cli\u003eDetails a wide range of electrophysiological techniques and spectroscopic methods used to analyze the function of ion channels, ion pumps and transporters\u003c\/li\u003e \u003cli\u003eCovers state-of-the art analytical methods to study ion pumps, channels, and transporters, and where analytical chemistry can make further contributions\u003c\/li\u003e \u003c\/ul\u003e\u003cbr\u003e\u003cbr\u003e\u003cb\u003eTrade Review\u003c\/b\u003e\u003cbr\u003e\"Overall \u003ci\u003ePumps, channels and transporters: methods of functional analysis\u003c\/i\u003e is an excellent book full of useful, detailed information and well worth reading whether you are an experienced cellular biologist or just a curious science undergraduate.\" (Chemistry in Australia 2016)\u003cbr\u003e\u003cbr\u003e\u003cb\u003eTable of Contents\u003c\/b\u003e\u003cbr\u003e\u003cp\u003ePreface xv\u003c\/p\u003e \u003cp\u003eList of Contributors xix\u003c\/p\u003e \u003cp\u003e\u003cb\u003e1 Introduction 1\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eMohammed A. A. Khalid and Ronald J. Clarke\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e1.1 History 1\u003c\/p\u003e \u003cp\u003e1.2 Energetics of Transport 6\u003c\/p\u003e \u003cp\u003e1.3 Mechanistic Considerations 7\u003c\/p\u003e \u003cp\u003e1.4 Ion Channels 8\u003c\/p\u003e \u003cp\u003e1.4.1 Voltage-Gated 8\u003c\/p\u003e \u003cp\u003e1.4.2 Ligand-Gated 9\u003c\/p\u003e \u003cp\u003e1.4.3 Mechanosensitive 9\u003c\/p\u003e \u003cp\u003e1.4.4 Light-Gated 9\u003c\/p\u003e \u003cp\u003e1.5 Ion Pumps 10\u003c\/p\u003e \u003cp\u003e1.5.1 ATP-Activated 10\u003c\/p\u003e \u003cp\u003e1.5.2 Light-Activated 11\u003c\/p\u003e \u003cp\u003e1.5.3 Redox-Linked 12\u003c\/p\u003e \u003cp\u003e1.6 Transporters 13\u003c\/p\u003e \u003cp\u003e1.6.1 Symporters and Antiporters 13\u003c\/p\u003e \u003cp\u003e1.6.2 Na+-Linked and H+-Linked 14\u003c\/p\u003e \u003cp\u003e1.7 Diseases of Ion Channels, Pumps, and Transporters 15\u003c\/p\u003e \u003cp\u003e1.7.1 Channelopathies 15\u003c\/p\u003e \u003cp\u003e1.7.2 Pump Dysfunction 17\u003c\/p\u003e \u003cp\u003e1.7.3 Transporter Dysfunction 18\u003c\/p\u003e \u003cp\u003e1.8 Conclusion 18\u003c\/p\u003e \u003cp\u003eReferences 19\u003c\/p\u003e \u003cp\u003e\u003cb\u003e2 Study of Ion Pump Activity Using Black Lipid Membranes 23\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eHans\u003c\/i\u003e\u003ci\u003e-\u003c\/i\u003e\u003ci\u003eJ\u003c\/i\u003e\u003ci\u003eü\u003c\/i\u003e\u003ci\u003ergen Apell and Valerij S. Sokolov\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e2.1 Introduction 23\u003c\/p\u003e \u003cp\u003e2.2 Formation of Black Lipid Membranes 24\u003c\/p\u003e \u003cp\u003e2.3 Reconstitution in Black Lipid Membranes 25\u003c\/p\u003e \u003cp\u003e2.3.1 Reconstitution of Na+,K+-ATPase in Black Lipid Membranes 25\u003c\/p\u003e \u003cp\u003e2.3.2 Recording Transient Currents with Membrane Fragments Adsorbed to a Black Lipid Membrane 26\u003c\/p\u003e \u003cp\u003e2.4 The Principles of Capacitive Coupling 28\u003c\/p\u003e \u003cp\u003e2.4.1 Dielectric Coefficients 29\u003c\/p\u003e \u003cp\u003e2.5 The Gated-Channel Concept 31\u003c\/p\u003e \u003cp\u003e2.6 Relaxation Techniques 34\u003c\/p\u003e \u003cp\u003e2.6.1 Concentration-Jump Methods 34\u003c\/p\u003e \u003cp\u003e2.6.2 Charge-Pulse Method 39\u003c\/p\u003e \u003cp\u003e2.7 Admittance Measurements 39\u003c\/p\u003e \u003cp\u003e2.8 The Investigation of Cytoplasmic and Extracellular Ion Access Channels in the Na+,K+-ATPase 42\u003c\/p\u003e \u003cp\u003e2.9 Conclusions 43\u003c\/p\u003e \u003cp\u003eReferences 45\u003c\/p\u003e \u003cp\u003e\u003cb\u003e3 Analyzing Ion Permeation in Channels and Pumps Using Patch\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eClamp Recording 51\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eAndrew J. Moorhouse, Trevor M. Lewis, and Peter H. Barry\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e3.1 Introduction 51\u003c\/p\u003e \u003cp\u003e3.2 Description of the Patch-Clamp Technique 52\u003c\/p\u003e \u003cp\u003e3.2.1 Development of Whole-Cell Dialysis with Voltage-Clamp 52\u003c\/p\u003e \u003cp\u003e3.3 Patch-Clamp Measurement and Analysis of Single Channel Conductance 54\u003c\/p\u003e \u003cp\u003e3.3.1 Conductance and Ohm’s Law 54\u003c\/p\u003e \u003cp\u003e3.3.2 Conductance of Channels versus Pumps 56\u003c\/p\u003e \u003cp\u003e3.3.3 Fluctuation Analysis 57\u003c\/p\u003e \u003cp\u003e3.3.4 Single Channel Recordings 61\u003c\/p\u003e \u003cp\u003e3.4 Determining Ion Selectivity and Relative Permeation in Whole-Cell Recordings 67\u003c\/p\u003e \u003cp\u003e3.4.1 Dilution Potential Measurements 67\u003c\/p\u003e \u003cp\u003e3.4.2 Bi-Ionic Potential Measurements 69\u003c\/p\u003e \u003cp\u003e3.4.3 Voltage and Solution Control in Whole-Cell Patch-Clamp Recordings 70\u003c\/p\u003e \u003cp\u003e3.4.4 Ion Shift Effects During Whole-Cell Patch-Clamp Experiments 71\u003c\/p\u003e \u003cp\u003e3.4.5 Liquid Junction Potential Corrections 72\u003c\/p\u003e \u003cp\u003e3.5 Influence of Voltage Corrections in Quantifying Ion Selectivity in Channels 74\u003c\/p\u003e \u003cp\u003e3.5.1 Analysis of Counterion Permeation in Glycine Receptor Channels 74\u003c\/p\u003e \u003cp\u003e3.5.2 Analysis of Anion-Cation Permeability in\u003c\/p\u003e \u003cp\u003eCation-Selective Mutant Glycine Receptor Channels 75\u003c\/p\u003e \u003cp\u003e3.6 Ion Permeation Pathways through Channels and Pumps 76\u003c\/p\u003e \u003cp\u003e3.6.1 The Ion Permeation Pathway in Pentameric Ligand-Gated Ion Channels 76\u003c\/p\u003e \u003cp\u003e3.6.1.1 Extracellular and Intracellular Components of the Permeation Pathway 78\u003c\/p\u003e \u003cp\u003e3.6.1.2 The TM2 Pore is the Primary Ion Selectivity Filter 79\u003c\/p\u003e \u003cp\u003e3.6.2 Ion Permeation Pathways in Pumps Identified Using Patch-Clamp 80\u003c\/p\u003e \u003cp\u003e3.6.2.1 Palytoxin Uncouples the Occluded Gates of the Na+,K+-ATPase 81\u003c\/p\u003e \u003cp\u003e3.7 Conclusions 82\u003c\/p\u003e \u003cp\u003eReferences 83\u003c\/p\u003e \u003cp\u003e\u003cb\u003e4 Probing Conformational Transitions of Membrane Proteins with Voltage Clamp Fluorometry (VCF) 89\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eThomas Friedrich\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e4.1 Introduction 89\u003c\/p\u003e \u003cp\u003e4.2 Description of The Vcf Technique 90\u003c\/p\u003e \u003cp\u003e4.2.1 Generation of Single-Cysteine Reporter Constructs, Expression in Xenopus laevis Oocytes, Site-Directed Fluorescence Labeling 90\u003c\/p\u003e \u003cp\u003e4.2.2 VCF Instrumentation 91\u003c\/p\u003e \u003cp\u003e4.2.3 Technical Precautions and Controls 93\u003c\/p\u003e \u003cp\u003e4.3 Perspectives from Early Measurements on Voltage-Gated K+ Channels 95\u003c\/p\u003e \u003cp\u003e4.3.1 Early Results Obtained with VCF on Voltage-Gated K+ Channels 95\u003c\/p\u003e \u003cp\u003e4.3.2 Probing the Environmental Changes: Fluorescence Spectra, Anisotropy, and the Effects of Quenchers 98\u003c\/p\u003e \u003cp\u003e4.4 Vcf Applied to P-Type Atpases 100\u003c\/p\u003e \u003cp\u003e4.4.1 Structural and Functional Aspects of Na+, K+- and H+,K+-ATPase 100\u003c\/p\u003e \u003cp\u003e4.4.2 The N790C Sensor Construct of Sheep Na+,K+-ATPase α1-Subunit 102\u003c\/p\u003e \u003cp\u003e4.4.2.1 Probing Voltage-Dependent Conformational Changes of Na+,K+-ATPase 103\u003c\/p\u003e \u003cp\u003e4.4.2.2 The Influence of Intracellular Na+ Concentrations 107\u003c\/p\u003e \u003cp\u003e4.4.3 The Rat Gastric H+,K+-ATPase S806C Sensor Construct 108\u003c\/p\u003e \u003cp\u003e4.4.3.1 Voltage-Dependent Conformational Shifts of the H+,K+-ATPase Sensor Construct S806C\u003c\/p\u003e \u003cp\u003eDuring the H+ Transport Branch 109\u003c\/p\u003e \u003cp\u003e4.4.3.2 An Intra- or Extracellular Access Channel of the Proton Pump? 110\u003c\/p\u003e \u003cp\u003e4.4.3.3 Effects of Extracellular Ligands: K+ and Na+ 111\u003c\/p\u003e \u003cp\u003e4.4.4 Probing Intramolecular Distances by Double Labeling and FRET 113\u003c\/p\u003e \u003cp\u003e4.5 Conclusions and Perspectives 116\u003c\/p\u003e \u003cp\u003eReferences 117\u003c\/p\u003e \u003cp\u003e\u003cb\u003e5 Patch Clamp Analysis of Transporters via Pre\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eSteady\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eState Kinetic Methods 121\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eChristof Grewer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e5.1 Introduction 121\u003c\/p\u003e \u003cp\u003e5.2 Patch Clamp Analysis of Secondary-Active Transporter Function 122\u003c\/p\u003e \u003cp\u003e5.2.1 Patch Clamp Methods 122\u003c\/p\u003e \u003cp\u003e5.2.2 Whole-Cell Recording 124\u003c\/p\u003e \u003cp\u003e5.2.3 Recording from Excised Patches 124\u003c\/p\u003e \u003cp\u003e5.3 Perturbation Methods 125\u003c\/p\u003e \u003cp\u003e5.3.1 Concentration Jumps 126\u003c\/p\u003e \u003cp\u003e5.3.2 Voltage Jumps 129\u003c\/p\u003e \u003cp\u003e5.4 Evaluation and Interpretation of Pre-Steady-State Kinetic Data 130\u003c\/p\u003e \u003cp\u003e5.4.1 Integrating Rate Equations that Describe Mechanistic Transport Models 131\u003c\/p\u003e \u003cp\u003e5.4.2 Assigning Kinetic Components to Elementary processes in the Transport Cycle 131\u003c\/p\u003e \u003cp\u003e5.5 Mechanistic Insight into Transporter Function 133\u003c\/p\u003e \u003cp\u003e5.5.1 Sequential Binding Mechanism 133\u003c\/p\u003e \u003cp\u003e5.5.2 Electrostatics 134\u003c\/p\u003e \u003cp\u003e5.5.3 Structure-Function Analysis 134\u003c\/p\u003e \u003cp\u003e5.6 Case Studies 136\u003c\/p\u003e \u003cp\u003e5.6.1 Glutamate Transporter Mechanism 136\u003c\/p\u003e \u003cp\u003e5.6.2 Electrogenic Charge Movements Associated with the Electroneutral Amino Acid Exchanger ASCT2 137\u003c\/p\u003e \u003cp\u003e5.7 Conclusions 139\u003c\/p\u003e \u003cp\u003eReferences 139\u003c\/p\u003e \u003cp\u003e\u003cb\u003e6 Recording of Pump and Transporter Activity Using Solid\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eSupported Membranes (SSM\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eBased Electrophysiology) 147\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eFrancesco Tadini\u003c\/i\u003e\u003ci\u003e-\u003c\/i\u003e\u003ci\u003eBuoninsegni and Klaus Fendler\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e6.1 Introduction 147\u003c\/p\u003e \u003cp\u003e6.2 The Instrument 148\u003c\/p\u003e \u003cp\u003e6.2.1 Rapid Solution Exchange Cuvette 149\u003c\/p\u003e \u003cp\u003e6.2.2 Setup and Flow Protocols 150\u003c\/p\u003e \u003cp\u003e6.2.3 Protein Preparations 151\u003c\/p\u003e \u003cp\u003e6.2.4 Commercial Instruments 152\u003c\/p\u003e \u003cp\u003e6.3 Measurement Procedures, Data Analysis, and Interpretation 152\u003c\/p\u003e \u003cp\u003e6.3.1 Current Measurement, Signal Analysis, and Reconstruction of Pump Currents 152\u003c\/p\u003e \u003cp\u003e6.3.2 Voltage Measurement 156\u003c\/p\u003e \u003cp\u003e6.3.3 Solution Exchange Artifacts 157\u003c\/p\u003e \u003cp\u003e6.4 P-Type Atp ases Investigated by Ssm-Based Electrophysiology 159\u003c\/p\u003e \u003cp\u003e6.4.1 Sarcoplasmic Reticulum Ca2+-ATPase 159\u003c\/p\u003e \u003cp\u003e6.4.2 Human Cu+-ATPases ATP7A and ATP7B 163\u003c\/p\u003e \u003cp\u003e6.5 Secondary Active Transporters 165\u003c\/p\u003e \u003cp\u003e6.5.1 Antiport: Assessing the Forward and Reverse Modes of the NhaA Na+\/H+ Exchanger of E. coli 166\u003c\/p\u003e \u003cp\u003e6.5.2 Cotransport: A Sugar-Induced Electrogenic Partial Reaction in the Lactose Permease LacY of E. coli 168\u003c\/p\u003e \u003cp\u003e6.5.3 The Glutamate Transporter EAAC1: A Robust Electrophysiological Assay with High Information Content 170\u003c\/p\u003e \u003cp\u003e6.6 Conclusions 172\u003c\/p\u003e \u003cp\u003eReferences 173\u003c\/p\u003e \u003cp\u003e\u003cb\u003e7 Stopped\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eFlow Fluorimetry Using Voltage\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eSensitive Fluorescent Membrane Probes 179\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eRonald J. Clarke and Mohammed A. A. Khalid\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e7.1 Introduction 179\u003c\/p\u003e \u003cp\u003e7.2 Basics of the Stopped-Flow Technique 181\u003c\/p\u003e \u003cp\u003e7.2.1 Flow Cell Design 181\u003c\/p\u003e \u003cp\u003e7.2.2 Rapid Data Acquisition 181\u003c\/p\u003e \u003cp\u003e7.2.3 Dead Time 183\u003c\/p\u003e \u003cp\u003e7.3 Covalent Versus Noncovalent Fluorescence Labeling 184\u003c\/p\u003e \u003cp\u003e7.3.1 Intrinsic Fluorescence 185\u003c\/p\u003e \u003cp\u003e7.3.2 Covalently Bound Extrinsic Fluorescent Probes 186\u003c\/p\u003e \u003cp\u003e7.3.3 Noncovalently Bound Extrinsic Fluorescent Probes 187\u003c\/p\u003e \u003cp\u003e7.4 Classes of Voltage-Sensitive Dyes 188\u003c\/p\u003e \u003cp\u003e7.4.1 Slow Dyes 188\u003c\/p\u003e \u003cp\u003e7.4.2 Fast Dyes 190\u003c\/p\u003e \u003cp\u003e7.5 Measurement of the Kinetics of the Na+,K+-Atpase 193\u003c\/p\u003e \u003cp\u003e7.5.1 Dye Concentration 194\u003c\/p\u003e \u003cp\u003e7.5.2 Excitation Wavelength and Light Source 197\u003c\/p\u003e \u003cp\u003e7.5.3 Monochromators and Filters 198\u003c\/p\u003e \u003cp\u003e7.5.4 Photomultiplier and Voltage Supply 199\u003c\/p\u003e \u003cp\u003e7.5.5 Reactions Detected by RH421 200\u003c\/p\u003e \u003cp\u003e7.5.6 Origin of the RH421 Response 202\u003c\/p\u003e \u003cp\u003e7.6 Conclusions 204\u003c\/p\u003e \u003cp\u003eReferences 204\u003c\/p\u003e \u003cp\u003e\u003cb\u003e8 Nuclear Magnetic Resonance Spectroscopy 211\u003cbr\u003e\u003c\/b\u003e\u003ci\u003ePhilip W. Kuchel\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e8.1 Introduction 211\u003c\/p\u003e \u003cp\u003e8.1.1 Definition of NMR 212\u003c\/p\u003e \u003cp\u003e8.1.2 Why So Useful? 212\u003c\/p\u003e \u003cp\u003e8.1.3 Magnetic Polarization 212\u003c\/p\u003e \u003cp\u003e8.1.4 Larmor Equation 213\u003c\/p\u003e \u003cp\u003e8.1.5 Chemical Shift 213\u003c\/p\u003e \u003cp\u003e8.1.6 Free Induction Decay 214\u003c\/p\u003e \u003cp\u003e8.1.7 Pulse Excitation 215\u003c\/p\u003e \u003cp\u003e8.1.8 Relaxation Times 217\u003c\/p\u003e \u003cp\u003e8.1.9 Splitting of Resonance Lines 217\u003c\/p\u003e \u003cp\u003e8.1.10 Measuring Membrane Transport 217\u003c\/p\u003e \u003cp\u003e8.2 Covalently-Induced Chemical Shift Differences 218\u003c\/p\u003e \u003cp\u003e8.2.1 Arginine Transport 218\u003c\/p\u003e \u003cp\u003e8.2.2 Other Examples 220\u003c\/p\u003e \u003cp\u003e8.3 Shift-Reagent-Induced Chemical Shift Differences 220\u003c\/p\u003e \u003cp\u003e8.3.1 DyPPP 220\u003c\/p\u003e \u003cp\u003e8.3.2 TmDTPA and TmDOTP 220\u003c\/p\u003e \u003cp\u003e8.3.3 Fast Cation Exchange 220\u003c\/p\u003e \u003cp\u003e8.4 pH-Induced Chemical Shift Differences 223\u003c\/p\u003e \u003cp\u003e8.4.1 Orthophosphate 223\u003c\/p\u003e \u003cp\u003e8.4.2 Methylphosphonate 224\u003c\/p\u003e \u003cp\u003e8.4.3 Triethylphosphate: 31P Shift Reference 224\u003c\/p\u003e \u003cp\u003e8.5 Hydrogen-Bond-Induced Chemical Shift Differences 225\u003c\/p\u003e \u003cp\u003e8.5.1 Phosphonates: DMMP 225\u003c\/p\u003e \u003cp\u003e8.5.2 HPA 225\u003c\/p\u003e \u003cp\u003e8.5.3 Fluorides 227\u003c\/p\u003e \u003cp\u003e8.6 Ionic-Environment-Induced Chemical Shift Differences 229\u003c\/p\u003e \u003cp\u003e8.6.1 Cs+ Transport 229\u003c\/p\u003e \u003cp\u003e8.7 Relaxation Time Differences 229\u003c\/p\u003e \u003cp\u003e8.7.1 Mn2+ Doping 229\u003c\/p\u003e \u003cp\u003e8.8 Diffusion Coefficient Differences 231\u003c\/p\u003e \u003cp\u003e8.8.1 Stejskal-Tanner Plot 231\u003c\/p\u003e \u003cp\u003e8.8.2 Andrasko’s Method 231\u003c\/p\u003e \u003cp\u003e8.9 Some Subtle Spectral Effects 233\u003c\/p\u003e \u003cp\u003e8.9.1 Scalar (J) Coupling Differences 233\u003c\/p\u003e \u003cp\u003e8.9.2 Endogenous Magnetic Field Gradients 233\u003c\/p\u003e \u003cp\u003e8.9.2.1 Magnetic Induction and Magnetic Field Strength 234\u003c\/p\u003e \u003cp\u003e8.9.2.2 Magnetic Field Gradients Across Cell Membranes and CO Treatment of RBCs 234\u003c\/p\u003e \u003cp\u003e8.9.2.3 Exploiting Magnetic Field Gradients in Membrane Transport Studies 235\u003c\/p\u003e \u003cp\u003e8.9.3 Residual Quadrupolar (νQ) Coupling 235\u003c\/p\u003e \u003cp\u003e8.10 A Case Study: The Stoichiometric Relationship Between the Number of Na+ Ions Transported per Molecule of Glucose Consumed in Human Rbcs 236\u003c\/p\u003e \u003cp\u003e8.11 Conclusions 239\u003c\/p\u003e \u003cp\u003eReferences 239\u003c\/p\u003e \u003cp\u003e\u003cb\u003e9 Time\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eResolved and Surface\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eEnhanced Infrared Spectroscopy 245\u003cbr\u003e\u003c\/b\u003eJoachim Heberle\u003c\/p\u003e \u003cp\u003e9.1 Introduction 245\u003c\/p\u003e \u003cp\u003e9.2 Basics of Ir Spectroscopy 246\u003c\/p\u003e \u003cp\u003e9.2.1 Vibrational Spectroscopy 246\u003c\/p\u003e \u003cp\u003e9.2.2 FTIR Spectroscopy 247\u003c\/p\u003e \u003cp\u003e9.2.3 IR Spectra of Biological Compounds 248\u003c\/p\u003e \u003cp\u003e9.2.4 Difference Spectroscopy 250\u003c\/p\u003e \u003cp\u003e9.3 Reflection Techniques 250\u003c\/p\u003e \u003cp\u003e9.3.1 Attenuated Total Reflection 250\u003c\/p\u003e \u003cp\u003e9.3.2 Surface-Enhanced IR Absorption 251\u003c\/p\u003e \u003cp\u003e9.4 Application to Electron-Transferring Proteins 252\u003c\/p\u003e \u003cp\u003e9.4.1 Cytochrome c 252\u003c\/p\u003e \u003cp\u003e9.4.2 Cytochrome c Oxidase 253\u003c\/p\u003e \u003cp\u003e9.5 Time-Resolved ir Spectroscopy 254\u003c\/p\u003e \u003cp\u003e9.5.1 The Rapid-Scan Technique 254\u003c\/p\u003e \u003cp\u003e9.5.2 The Step-Scan Technique 255\u003c\/p\u003e \u003cp\u003e9.5.3 Tunable QCLs 255\u003c\/p\u003e \u003cp\u003e9.6 Applications to Retinal Proteins 256\u003c\/p\u003e \u003cp\u003e9.6.1 Bacteriorhodopsin 256\u003c\/p\u003e \u003cp\u003e9.6.2 Channelrhodopsin 260\u003c\/p\u003e \u003cp\u003e9.7 Conclusions 263\u003c\/p\u003e \u003cp\u003eReferences 264\u003c\/p\u003e \u003cp\u003e10 Analysis of Membrane-Protein Complexes by Single-Molecule Methods 269\u003cbr\u003e\u003ci\u003eKatia Cosentino, Stephanie Bleicken, and Ana J. García\u003c\/i\u003e\u003ci\u003e-\u003c\/i\u003e\u003ci\u003eS\u003c\/i\u003e\u003ci\u003eá\u003c\/i\u003e\u003ci\u003eez\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e10.1 Introduction 269\u003c\/p\u003e \u003cp\u003e10.2 Fluorophores for Single Particle Labeling 270\u003c\/p\u003e \u003cp\u003e10.3 Principles of Fluorescence Correlation Spectroscopy 271\u003c\/p\u003e \u003cp\u003e10.3.1 Analysis of Molecular Complexes by Two-Color FCS 275\u003c\/p\u003e \u003cp\u003e10.3.2 FCS Variants to Study Lipid Membranes 275\u003c\/p\u003e \u003cp\u003e10.3.3 FCS Applications to Membranes 278\u003c\/p\u003e \u003cp\u003e10.4 Principle and Analysis of Single-Molecule Imaging 279\u003c\/p\u003e \u003cp\u003e10.4.1 TIRF Microscopy 280\u003c\/p\u003e \u003cp\u003e10.4.2 Single-Molecule Detection 282\u003c\/p\u003e \u003cp\u003e10.4.3 Single Particle Tracking and Trajectory Analysis 284\u003c\/p\u003e \u003cp\u003e10.5 Complex Dynamics and Stoichiometry by Single-Molecule Microscopy 285\u003c\/p\u003e \u003cp\u003e10.5.1 Application to Single-Molecule Stoichiometry Analysis 285\u003c\/p\u003e \u003cp\u003e10.5.2 Application to Kinetics Processes in Cell Membranes 290\u003c\/p\u003e \u003cp\u003e10.6 Fcs Versus Spt 291\u003c\/p\u003e \u003cp\u003eReferences 291\u003c\/p\u003e \u003cp\u003e\u003cb\u003e11 Probing Channel, Pump, and Transporter Function Using Single\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eMolecule Fluorescence 299\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eEve E. Weatherill, John S. H. Danial, and Mark I. Wallace\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e11.1 Introduction 299\u003c\/p\u003e \u003cp\u003e11.1.1 Basic Principles 300\u003c\/p\u003e \u003cp\u003e11.2 Practical Considerations 300\u003c\/p\u003e \u003cp\u003e11.2.1 Observables 301\u003c\/p\u003e \u003cp\u003e11.2.2 Apparatus 301\u003c\/p\u003e \u003cp\u003e11.2.3 Labels 302\u003c\/p\u003e \u003cp\u003e11.2.4 Bilayers 303\u003c\/p\u003e \u003cp\u003e11.3 smf Imaging 303\u003c\/p\u003e \u003cp\u003e11.3.1 Fluorescence Colocalization 304\u003c\/p\u003e \u003cp\u003e11.3.2 Conformational Changes 306\u003c\/p\u003e \u003cp\u003e11.3.3 Superresolution Microscopy 307\u003c\/p\u003e \u003cp\u003e11.4 Single Molecule Förster Resonance Energy Transfer 308\u003c\/p\u003e \u003cp\u003e11.4.1 Interactions\/Stoichiometry 308\u003c\/p\u003e \u003cp\u003e11.4.2 Conformational Changes 309\u003c\/p\u003e \u003cp\u003e11.5 Single-Molecule Counting by Photobleaching 312\u003c\/p\u003e \u003cp\u003e11.6 Optical Channel Recording 314\u003c\/p\u003e \u003cp\u003e11.7 Simultaneous Techniques 315\u003c\/p\u003e \u003cp\u003e11.8 Summary 318\u003c\/p\u003e \u003cp\u003eReferences 318\u003c\/p\u003e \u003cp\u003e\u003cb\u003e12 Electron Paramagnetic Resonance: Site\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eDirected Spin Labeling 327\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eLouise J. Brown and Joanna E. Hare\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e12.1 Introduction 327\u003c\/p\u003e \u003cp\u003e12.1.1 Development of EPR as a Tool for Structural Biology 329\u003c\/p\u003e \u003cp\u003e12.1.2 SDSL-EPR: A Complementary Approach to Determine Structure-Function Relationships 330\u003c\/p\u003e \u003cp\u003e12.2 Basics of the Epr Method 331\u003c\/p\u003e \u003cp\u003e12.2.1 Physical Basis of the EPR Signal 331\u003c\/p\u003e \u003cp\u003e12.2.2 Spin Labeling 333\u003c\/p\u003e \u003cp\u003e12.2.3 EPR Instrumentation 336\u003c\/p\u003e \u003cp\u003e12.3 Structural and Dynamic Information from Sdsl-Epr 336\u003c\/p\u003e \u003cp\u003e12.3.1 Mobility Measurements 336\u003c\/p\u003e \u003cp\u003e12.3.2 Solvent Accessibility 341\u003c\/p\u003e \u003cp\u003e12.4 Distance Measurements 345\u003c\/p\u003e \u003cp\u003e12.4.1 Interspin Distance Measurements 345\u003c\/p\u003e \u003cp\u003e12.4.2 Continuous Wave 347\u003c\/p\u003e \u003cp\u003e12.4.3 Pulsed Methods: DEER 349\u003c\/p\u003e \u003cp\u003e12.5 Challenges 353\u003c\/p\u003e \u003cp\u003e12.5.1 New Labels 353\u003c\/p\u003e \u003cp\u003e12.5.2 Spin-Label Flexibility 355\u003c\/p\u003e \u003cp\u003e12.5.3 Production and Reconstitution Challenges: Nanodiscs 355\u003c\/p\u003e \u003cp\u003e12.6 Conclusions 356\u003c\/p\u003e \u003cp\u003eReferences 357\u003c\/p\u003e \u003cp\u003e\u003cb\u003e13 Radioactivity\u003c\/b\u003e\u003cb\u003e-\u003c\/b\u003e\u003cb\u003eBased Analysis of Ion Transport 367\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eIngolf Bernhardt and J. Clive Ellory\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e13.1 Introduction 367\u003c\/p\u003e \u003cp\u003e13.2 Membrane Permeability for Electroneutral Substances and Ions 368\u003c\/p\u003e \u003cp\u003e13.3 Kinetic Considerations 370\u003c\/p\u003e \u003cp\u003e13.4 Techniques for Ion Flux Measurements 371\u003c\/p\u003e \u003cp\u003e13.4.1 Conventional Methods 371\u003c\/p\u003e \u003cp\u003e13.4.2 Alternative Method 373\u003c\/p\u003e \u003cp\u003e13.5 Kinetic Analysis of Ion Transporter Properties 375\u003c\/p\u003e \u003cp\u003e13.6 Selected Cation Transporter Studies on Red Blood Cells 376\u003c\/p\u003e \u003cp\u003e13.6.1 K+,Cl− Cotransport (KCC) 378\u003c\/p\u003e \u003cp\u003e13.6.2 Residual Transport 378\u003c\/p\u003e \u003cp\u003e13.7 Combination of Radioactive Isotope Studies with Methods using Fluorescent Dyes 379\u003c\/p\u003e \u003cp\u003e13.8 Conclusions 382\u003c\/p\u003e \u003cp\u003eReferences 383\u003c\/p\u003e \u003cp\u003e\u003cb\u003e14 Cation Uptake Studies with Atomic Absorption Spectrophotometry (Aas) 387\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eThomas Friedrich\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e14.1 Introduction 387\u003c\/p\u003e \u003cp\u003e14.2 Overview of the Technique of Aas 389\u003c\/p\u003e \u003cp\u003e14.2.1 Historical Account of AAS with Flame Atomization 390\u003c\/p\u003e \u003cp\u003e14.2.2 Element-Specific Radiation Sources 391\u003c\/p\u003e \u003cp\u003e14.2.3 Electrothermal Atomization in Heated Graphite Tubes 392\u003c\/p\u003e \u003cp\u003e14.2.4 Correction for Background Absorption 394\u003c\/p\u003e \u003cp\u003e14.3 The Expression System of Xenopus laevis Oocytes for Cation Flux Studies: Practical Considerations 395\u003c\/p\u003e \u003cp\u003e14.4 Experimental Outline of the Aas Flux Quantification Technique 395\u003c\/p\u003e \u003cp\u003e14.5 Representative Results Obtained with the Aas Flux Quantification Technique 397\u003c\/p\u003e \u003cp\u003e14.5.1 Reaction Cycle of P-Type ATPases 398\u003c\/p\u003e \u003cp\u003e14.5.2 Rb+ Uptake Kinetics: Inhibitor Sensitivity 398\u003c\/p\u003e \u003cp\u003e14.5.3 Dependence of Rb+ Transport of Gastric H+,K+-ATPase on Extra- and Intracellular pH 400\u003c\/p\u003e \u003cp\u003e14.5.4 Determination of Na+,K+-ATPase Transport Stoichiometry and Voltage Dependence of H+,K+-ATPase Rb+ Transport 403\u003c\/p\u003e \u003cp\u003e14.5.5 Effects of C-Terminal Deletions of the H+,K+-ATPase α-Subunit 404\u003c\/p\u003e \u003cp\u003e14.5.6 Li+ and Cs+ Uptake Studies 405\u003c\/p\u003e \u003cp\u003e14.6 Concluding Remarks 407\u003c\/p\u003e \u003cp\u003eReferences 408\u003c\/p\u003e \u003cp\u003e\u003cb\u003e15 Long Timescale Molecular Simulations for Understanding Ion Channel Function 411\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eBen Corry\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e15.1 Introduction 411\u003c\/p\u003e \u003cp\u003e15.2 Fundamentals of Md Simulation 412\u003c\/p\u003e \u003cp\u003e15.2.1 The Main Idea 412\u003c\/p\u003e \u003cp\u003e15.2.2 Force Fields 414\u003c\/p\u003e \u003cp\u003e15.2.3 O ther Simulation Considerations 416\u003c\/p\u003e \u003cp\u003e15.2.4 Why Do MD Simulations Take So Much Computational Power? 416\u003c\/p\u003e \u003cp\u003e15.2.4.1 Force Calculations 417\u003c\/p\u003e \u003cp\u003e15.2.4.2 Time Step 417\u003c\/p\u003e \u003cp\u003e15.3 Simulation Duration and Simulation Size 418\u003c\/p\u003e \u003cp\u003e15.4 Historical Development of Long Md Simulations 421\u003c\/p\u003e \u003cp\u003e15.5 Limitations and Challenges Facing Md Simulations 423\u003c\/p\u003e \u003cp\u003e15.5.1 Force Field and Algorithm Accuracy 423\u003c\/p\u003e \u003cp\u003e15.5.2 Sampling Problems 424\u003c\/p\u003e \u003cp\u003e15.6 Example Simulations of Ion Channels 425\u003c\/p\u003e \u003cp\u003e15.6.1 Simulations of Ion Permeation 425\u003c\/p\u003e \u003cp\u003e15.6.2 Simulations of Ion Selectivity 428\u003c\/p\u003e \u003cp\u003e15.6.3 Simulations of Channel Gating 432\u003c\/p\u003e \u003cp\u003e15.7 Conclusions 433\u003c\/p\u003e \u003cp\u003eReferences 436\u003c\/p\u003e \u003cp\u003eIndex 443\u003c\/p\u003e \u003cp\u003eChemical Analysis: A Series of Monographs on Analytical\u003c\/p\u003e \u003cp\u003eChemistry and its Applications 461\u003c\/p\u003e","brand":"John Wiley \u0026 Sons Inc","offers":[{"title":"Default Title","offer_id":49406936252759,"sku":"9781118858806","price":97.16,"currency_code":"GBP","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0817\/1739\/5799\/files\/9781118858806.jpg?v=1730497614","url":"https:\/\/bookcurl.com\/products\/pumps-channels-and-transporters-9781118858806","provider":"Book Curl","version":"1.0","type":"link"}