Metabolic Analysis Using Stable Isotopes

1st Edition - September 8, 2015

Editor: Christian Metallo

Hardback ISBN:

9 7 8 - 0 - 1 2 - 8 0 2 2 9 3 - 1

eBook ISBN:

9 7 8 - 0 - 1 2 - 8 0 2 5 3 4 - 5

Metabolic Analysis Using Stable Isotopes, the newest volume in Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the… Read more

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Metabolic Analysis Using Stable Isotopes

, the newest volume in Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in metabolic analysis using stable isotopes.

Dedication
Preface
Chapter One: Studies of Metabolism Using ¹³C MRS of Hyperpolarized Probes
- Abstract
- 1 Introduction
- 2 ¹³C MRS of DNP Probes: Current Approaches to Studying Metabolism
- 3 Requirements of ¹³C DNP Probes
- 4 Conducting Hyperpolarized Experiments
- 5 Acquisition Strategies for Hyperpolarized MR Spectroscopy and Imaging
- 6 Analysis of Hyperpolarized MR Data
- 7 Summary and Conclusion
Chapter Two: Hyperpolarized ¹³C Magnetic Resonance and Its Use in Metabolic Assessment of Cultured Cells and Perfused Organs
- Abstract
- 1 Introduction: Importance of Developing Methods to Observe Metabolic Flux in Disease States
- 2 Hyperpolarization Methods and Sample Preparation
- 3 Dynamic Assessment of Metabolism in Cells
- 4 Dynamic Assessment of Metabolism in Perfused Organs
- 5 Challenges and Future Directions
- 6 Conclusion
- Acknowledgments
Chapter Three: Metabolic Tracing Using Stable Isotope-Labeled Substrates and Mass Spectrometry in the Perfused Mouse Heart
- Abstract
- 1 Introduction
- 2 Study Model: Mouse Heart Perfusion Ex Vivo in the Working Mode with Semi-Recirculating Buffer
- 3 Metabolic Flux Measurements in the Perfused Mouse Heart Using ¹³C-Labeled Substrates and Gas Chromatography-Mass Spectrometry
- 4 Practical Considerations and Future Challenges
- Acknowledgments
Chapter Four: Probing Metabolism in the Intact Retina Using Stable Isotope Tracers
- Abstract
- 1 Introduction
- 2 Methods
- 3 Applications
- 4 Summary
- Acknowledgments
Chapter Five: Analysis of Cell Metabolism Using LC-MS and Isotope Tracers
- Abstract
- 1 Introduction
- 2 Methods for Sample Extraction
- 3 LC-MS Methods
- 4 LC-MS Data Analysis
- 5 Use of Stable Isotope Tracers
- 6 Normalization
- 7 Quantification of Metabolites in Cells and Medium
- 8 Stable Isotope Tracing with Fumarate Hydratase-Deficient Cell Model
- 9 Future
Chapter Six: Analysis of Fatty Acid Metabolism Using Stable Isotope Tracers and Mass Spectrometry
- Abstract
- 1 Introduction
- 2 Labeling and Extraction of Fatty Acids in Cultured Cells
- 3 Mass Spectrometry Analysis of Fatty Acid Labeling
- 4 Fatty Acid-Labeling Data Analysis and Interpretation
- 5 Summary and Perspective
Chapter Seven: Dynamic Proteomics: In Vivo Proteome-Wide Measurement of Protein Kinetics Using Metabolic Labeling
- Abstract
- 1 Introduction
- 2 Fundamentals: Stable Isotope-Labeling Approaches for Measuring Protein Synthesis and Other Polymerization Biosynthesis Rates
- 3 Application of Combinatorial Analysis (MIDA) Approaches to Protein Dynamics
- 4 Sample Processing
- 5 Analytical Methods
- 6 Future Directions
- Acknowledgments
Chapter Eight: Non-targeted Tracer Fate Detection
- Abstract
- 1 Introduction
- 2 Theoretical Background
- 3 Practical Considerations
- 4 NTFD Applications
- 5 Summary and Outlook
- Acknowledgment
Chapter Nine: Isotopomer Spectral Analysis: Utilizing Nonlinear Models in Isotopic Flux Studies
- Abstract
- 1 Introduction
- 2 Comparing Radioisotopes and Stable Isotopes
- 3 ISA Methodology
- 4 Linear Versus Nonlinear Models
- 5 Estimating ISA Parameter Values Using Nonlinear Regression
- 6 Examples of ISA Models
- 7 Concluding Thoughts
Chapter Ten: Effect of Error Propagation in Stable Isotope Tracer Studies: An Approach for Estimating Impact on Apparent Biochemical Flux
- Abstract
- 1 Introduction
- 2 Analytical: Instrument Settings and Data Acquisition
- 3 Simulations of Lipid Flux
- 4 Simulations of Protein Flux
- 5 Summary and Conclusions
Author Index
Subject Index