Beschreibung
Inhaltsangabe13C-NMR in Metabolic Studies.- 1 Introduction.- 2 The Design and Interpretation of Labelling Experiments.- 3 Radioisotope Labelling.- 4 13C NMR Spectroscopy.- 4.1 Assignment of 13C NMR Spectra.- 4.1.1 Known Chemical Shifts and Substituent Chemical Shift Effects.- 4.1.2 Off-Resonance and Specific Proton Decoupling.- 4.1.3 Analysis of Long-Range 1H-13C Couplings.- 4.1.4 Polarisation Transfer and 2D NMR Methods.- 4.1.5 Lanthanide-Induced Shift Studies.- 4.1.6 Model and Derivative Studies.- 4.1.7 Synthesis of Isotopically Labelled Compounds.- 4.1.8 Incorporation Studies.- 5 Biosynthetic Methodology.- 5.1 Precursor Incorporation.- 6 13C Enrichment Studies.- 6.1 Single 13C-Labelling.- 6.1.1 Quantitative 13C Measurements.- 6.1.2 Single 13C-Labelling Studies in Plants.- 6.2 13C-13C Spin-Spin Coupling.- 6.2.1 Singly 13C-Labelled Precursors.- 6.2.2 Double 13C-Labelled Precursors.- 6.2.3 Non-Contiguous Double 13C-Labelling.- 6.2.3.1 Two-Bond 13C-13C Couplings from Contiguous 13C2-Labelled Precursors.- 6.2.3.2 One-Bond 13C-13C Couplings from Non-Contiguous 13C2-Labelled Precursors.- 6.2.3.3 Two-Bond Couplings from Non-Contiguous 13C2-Labelled Precursors.- 7 13C, 15N Doubly Labelled Precursors.- 8 Isotope-Induced Shifts in 13C NMR.- 8.1 The Deuterium Alpha-Shift Technique.- 8.2 The Deuterium Beta-Shift Technique.- 8.3 18O Isotope Induced Shifts in 13C NMR.- 9 Conclusion.- References.- Determination of the Energy Status of Plant Cells by 31P-Nuclear Magnetic Resonance Spectroscopy.- 1 Introduction.- 2 Principles.- 3 Experimental Considerations.- 4 Quantitation of Metabolite Concentrations of vivo by NMR.- 5 Measurement of Nucleoside Di- and Triphosphate Levels by 31P-NMR.- 6 Estimation of the Extent of Magnesium Ion Binding to Nucleoside Triphosphates in vivo by 31P-NMR.- 7 Measurement of the Gibbs' Free Energy for Hydrolysis of ATP.- 8 Relationship of in vivo 31P-NMR Measurements to What Is Seen by Other Methods.- 9 Conclusions.- References.- The Use of NMR Spectroscopy to Follow Deuterium in Studies of Fungal Metabolism.- 1 Introduction.- 2 Deuterium NMR Spectroscopy.- 2.1 Background and Techniques.- 2.1.1 The Nuclear Magnetic Properties of Deuterium.- 2.1.2 The Deuterium NMR Spectrum.- 2.1.3 Recording a Deuterium NMR Spectrum.- 2.2 Applications of Deuterium NMR Spectroscopy to Fungal Metabolism.- 2.2.1 Administering the Precursor.- 2.2.2 Establishing the Retention or Loss of a Hydrogen.- 2.2.3 Establishing Precursor-Product Relationships.- 2.2.4 Mechanistic Studies.- 2.2.4.1 Displacement Reactions.- 2.2.4.2 Incorporation of Deuterium Atoms from the Medium.- 2.2.4.3 Following Hydride Shifts.- 2.2.5 The Use of [2-2H3]-Acetate as a Polyketide Precursor.- 2.3 Kinetic Isotope Effects.- 2.4 The Use of Shift Reagents.- 3 Detection of Deuterium Indirectly in the 13C-NMR Spectrum.- 3.1 Background and Techniques.- 3.1.1 Detecting Deuterium Directly Attached to a Carbon Reporter Nucleus (the ?-Shift Technique).- 3.1.2 Detecting Deuterium on a Carbon Adjacent to the Reporter Nucleus (the ?-Shift Technique).- 3.2 Applications to the Study of Biosynthesis in Fungi.- 3.2.1 Quantitative Measurements of Deuterium Retention.- 3.2.2 Determining the Multiplicity of Labelling.- 3.2.3 Labelling a Bond.- 4 Conclusions.- References.- Proton NMR Studies on DNA Structure.- 1 Introduction.- 2 Theoretical Background.- 2.1 Chemical Shifts.- 2.2 Coupling Constants.- 2.3 NOE.- 2.4 Assignments.- 3 Examples.- 4 Summary.- References.- NMR Methods for Determination of Intracellular pH.- 1 Introduction.- 2 Some Principles.- 3 Determinants of Chemical Shift.- 4 Measurement of Chemical Shifts.- 5 Estimating Intracellular pH from Chemical Shifts: Titration Curves.- 6 Other Factors Contributing to Potential Errors in Intracellular pH Measurements Using NMR.- 7 Observation of pH Heterogeneity in Cell Populations and Tissues by NMR.- 8 NMR Signals That Have Been Used as Intracellular pH Indicators.- 9 Assignment of Resonances.- 10 Can the pH in Compartments Other Than Cytoplasm
Autorenporträt
Inhaltsangabe13C-NMR in Metabolic Studies.- 1 Introduction.- 2 The Design and Interpretation of Labelling Experiments.- 3 Radioisotope Labelling.- 4 13C NMR Spectroscopy.- 4.1 Assignment of 13C NMR Spectra.- 4.1.1 Known Chemical Shifts and Substituent Chemical Shift Effects.- 4.1.2 Off-Resonance and Specific Proton Decoupling.- 4.1.3 Analysis of Long-Range 1H-13C Couplings.- 4.1.4 Polarisation Transfer and 2D NMR Methods.- 4.1.5 Lanthanide-Induced Shift Studies.- 4.1.6 Model and Derivative Studies.- 4.1.7 Synthesis of Isotopically Labelled Compounds.- 4.1.8 Incorporation Studies.- 5 Biosynthetic Methodology.- 5.1 Precursor Incorporation.- 6 13C Enrichment Studies.- 6.1 Single 13C-Labelling.- 6.1.1 Quantitative 13C Measurements.- 6.1.2 Single 13C-Labelling Studies in Plants.- 6.2 13C-13C Spin-Spin Coupling.- 6.2.1 Singly 13C-Labelled Precursors.- 6.2.2 Double 13C-Labelled Precursors.- 6.2.3 Non-Contiguous Double 13C-Labelling.- 6.2.3.1 Two-Bond 13C-13C Couplings from Contiguous 13C2-Labelled Precursors.- 6.2.3.2 One-Bond 13C-13C Couplings from Non-Contiguous 13C2-Labelled Precursors.- 6.2.3.3 Two-Bond Couplings from Non-Contiguous 13C2-Labelled Precursors.- 7 13C, 15N Doubly Labelled Precursors.- 8 Isotope-Induced Shifts in 13C NMR.- 8.1 The Deuterium Alpha-Shift Technique.- 8.2 The Deuterium Beta-Shift Technique.- 8.3 18O Isotope Induced Shifts in 13C NMR.- 9 Conclusion.- References.- Determination of the Energy Status of Plant Cells by 31P-Nuclear Magnetic Resonance Spectroscopy.- 1 Introduction.- 2 Principles.- 3 Experimental Considerations.- 4 Quantitation of Metabolite Concentrations of vivo by NMR.- 5 Measurement of Nucleoside Di- and Triphosphate Levels by 31P-NMR.- 6 Estimation of the Extent of Magnesium Ion Binding to Nucleoside Triphosphates in vivo by 31P-NMR.- 7 Measurement of the Gibbs' Free Energy for Hydrolysis of ATP.- 8 Relationship of in vivo 31P-NMR Measurements to What Is Seen by Other Methods.- 9 Conclusions.- References.- The Use of NMR Spectroscopy to Follow Deuterium in Studies of Fungal Metabolism.- 1 Introduction.- 2 Deuterium NMR Spectroscopy.- 2.1 Background and Techniques.- 2.1.1 The Nuclear Magnetic Properties of Deuterium.- 2.1.2 The Deuterium NMR Spectrum.- 2.1.3 Recording a Deuterium NMR Spectrum.- 2.2 Applications of Deuterium NMR Spectroscopy to Fungal Metabolism.- 2.2.1 Administering the Precursor.- 2.2.2 Establishing the Retention or Loss of a Hydrogen.- 2.2.3 Establishing Precursor-Product Relationships.- 2.2.4 Mechanistic Studies.- 2.2.4.1 Displacement Reactions.- 2.2.4.2 Incorporation of Deuterium Atoms from the Medium.- 2.2.4.3 Following Hydride Shifts.- 2.2.5 The Use of [2-2H3]-Acetate as a Polyketide Precursor.- 2.3 Kinetic Isotope Effects.- 2.4 The Use of Shift Reagents.- 3 Detection of Deuterium Indirectly in the 13C-NMR Spectrum.- 3.1 Background and Techniques.- 3.1.1 Detecting Deuterium Directly Attached to a Carbon Reporter Nucleus (the ?-Shift Technique).- 3.1.2 Detecting Deuterium on a Carbon Adjacent to the Reporter Nucleus (the ?-Shift Technique).- 3.2 Applications to the Study of Biosynthesis in Fungi.- 3.2.1 Quantitative Measurements of Deuterium Retention.- 3.2.2 Determining the Multiplicity of Labelling.- 3.2.3 Labelling a Bond.- 4 Conclusions.- References.- Proton NMR Studies on DNA Structure.- 1 Introduction.- 2 Theoretical Background.- 2.1 Chemical Shifts.- 2.2 Coupling Constants.- 2.3 NOE.- 2.4 Assignments.- 3 Examples.- 4 Summary.- References.- NMR Methods for Determination of Intracellular pH.- 1 Introduction.- 2 Some Principles.- 3 Determinants of Chemical Shift.- 4 Measurement of Chemical Shifts.- 5 Estimating Intracellular pH from Chemical Shifts: Titration Curves.- 6 Other Factors Contributing to Potential Errors in Intracellular pH Measurements Using NMR.- 7 Observation of pH Heterogeneity in Cell Populations and Tissues by NMR.- 8 NMR Signals That Have Been Used as Intracellular pH Indicators.- 9 Assignment of Resonances.- 10 Can the pH in Compartments Other Than Cytoplasm
