Isotope labeling of biomolecules : applications /

Isotope Labeling of Biomolecules: Applications, the latest in the Methods in Enzymology series, focuses on stable isotope labeling methods and applications for biomolecules. This practical guide to biomolecular labeling looks at new techniques that are becoming widely used.

Detalles Bibliográficos
Clasificación:Libro Electrónico
Otros Autores: Kelman, Zvi (Editor )
Formato: Electrónico eBook
Idioma:Inglés
Publicado: Cambridge, MA : Academic Press, 2016.
Edición:First edition.
Colección:Methods in enzymology ; v. 566.
Temas:
Radiolabeling.
Biomolecules.
Isotope Labeling
Radiomarquage.
Biomol�ecules.
MEDICAL > Pharmacology.
Biomolecules
Radiolabeling
Internet Resources.
Acceso en línea:Texto completo
Texto completo
Texto completo
Tabla de Contenidos:
  • Front Cover
  • Isotope Labeling of Biomolecules
  • Applications
  • Copyright
  • Contents
  • Contributors
  • Preface
  • Section I: Nuclear Magnetic Resonance Spectroscopy
  • Chapter One: Application of Natural Isotopic Abundance 1H-13C- and 1H-15N-Correlated Two-Dimensional NMR for Evaluation o ...
  • 1. Introduction
  • 2. Preparation of Protein Therapeutic Samples
  • 2.1. Considerations for Sample Conditions
  • 2.2. Sample Preparation of the Intact NISTmAb
  • 2.3. Enzymatic Digest of mAbs into Fab and Fc Domains
  • 2.4. Sample Preparation of Filgrastim
  • 3. Acquisition of 2D Spectra.
  • 3.1. Amide 1H-15N-Correlated Spectra
  • 3.2. Methyl 1H-13C-Correlated Spectra
  • 3.3. Nonuniform Sampling Method
  • 4. Processing 2D-Correlated Spectra
  • 4.1. Processing Uniformly Sampled Data
  • 4.2. Processing Nonuniformly Sampled Data
  • 5. Comparative Analysis of 2D-Correlated Spectra
  • 6. Perspectives
  • Acknowledgments
  • References
  • Chapter Two: A Semiautomated Assignment Protocol for Methyl Group Side Chains in Large Proteins
  • 1. Introduction
  • 2. Labeling of Side Chain Methyl Groups for Large Proteins
  • 3. Methyl Labeling Protocol for the cAMP-Dependent Protein Kinase A.
  • 3.1. Materials
  • 3.2. Purification of PKA-C
  • 3.3. Materials
  • 4. Semiautomated Methyl-Group Resonance Assignment Strategies
  • 5. Semiautomated Assignment Protocol Using FLAMEnGO 2.0
  • 5.1. Materials
  • 5.2. System Requirements and Specifications
  • 5.3. Input File Formats
  • 5.3.1. Initial Random Assignment
  • 5.3.2. 3D C,C,H 13C-HMQC-NOESY-13C-HMQC
  • 5.3.3. [1H,13C]-HMQC/15N Amide
  • 5.3.4. Paramagnetic Relaxation Enhancement Data
  • 5.3.5. Prediction of Methyl Group Chemical Shifts from the PDB File
  • 5.3.6. Residue Type
  • 5.4. Instructions to Run FLAMEnGO 2.0 GUI
  • 5.5. Output Files.
  • 5.5.1. Score and Assignment
  • 5.5.2. Summary
  • 6. Conclusions and Perspectives
  • Acknowledgments
  • References
  • Chapter Three: 19F-Site-Specific-Labeled Nucleotides for Nucleic Acid Structural Analysis by NMR
  • 1. Introduction
  • 2. General Enzymatic Synthesis of 2F-ATP, 5F-CTP, and 5F-UTP
  • 3. Stability of Modified RNA
  • 3.1. Impact of Fluorine Substitutions on Base Pairing
  • 3.2. Thermal Stability of Fluoro-Modified RNA
  • 4. Diagnostics of 19F-Chemical Shifts
  • 5. NMR-Assignment Strategies for Fluoro-Modified RNA
  • 5.1. General NMR Hardware Considerations
  • 5.2. 19F,19F-NOESY.
  • 5.3. 19F,1H-HOESY
  • 5.4. 1H,19F-HMBC
  • 6. Coupling Measurements in Fluoro-Modified RNA
  • 6.1. Scalar J-Coupling Measurements
  • 6.2. RDC Measurements
  • 7. Solvent-Induced Isotope Shifts
  • 8. Ligand-Induced Chemical Shift Changes
  • 9. Conclusion and Remarks
  • Acknowledgments
  • References
  • Chapter Four: Applying Thymine Isostere 2,4-Difluoro-5-Methylbenzene as a NMR Assignment Tool and Probe of Homopyrimidine...
  • 1. Introduction
  • 2. Preparation of dF-Substituted Oligonucleotides for NMR Analysis
  • 2.1. Design of Oligonucleotides
  • 2.2. Reagents.

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