Description

This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. The first of 2 volumes covering nucleosomes, histones and chromatin, it has chapters on methods applied to the study of protein arginine methylation, high-resolution identification of intra- and interchromosomal DNA interactions by 4C technology, and peptide arrays to interrogate the binding specificity of chromatin-binding proteins.

Key Features

  • Continues the legacy of this premier serial by containing quality chapters authored by leaders in the field
  • The first of 2 volumes covering nucleosomes, histones and chromatin
  • Chapters on methods applied to the study of protein arginine methylation, high-resolution identification of intra- and interchromosomal DNA interactions by 4C technology, and peptide arrays to interrogate the binding specificity of chromatin-binding proteins

Readership

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

Table of Contents

Series Page

Contributors

Preface

Volume in series

Chapter One Examining Histone Posttranslational Modification Patterns by High-Resolution Mass Spectrometry

1 Introduction

2 Materials

3 Nuclei Isolation

4 Acid Extraction

5 Offline Reverse-Phase High-Performance Liquid Chromatography for Histone Variant Purification (Optional)

6 Chemical Derivatization of Histones

7 Stage-Tip Cleanup of Peptide Samples Prior to MS Analysis

8 Enrichment for Phosphorylated Peptides Using Titanium Dioxide (TiO2) Beads (Optional)

9 Online RP-HPLC and MS

Chapter Two Investigating the Cell Cycle-Associated Dynamics of Histone Modifications Using Quantitative Mass Spectrometry

1 Introduction

2 Experimental Design

3 Cell Culture

4 Extraction and Separation of Histones

5 Preparation of Propionic Anhydride-Derivatized Tryptic Histone Peptides

6 Mass Spectrometric Analysis and Quantification of Histone PTMs

Chapter Three A Method to Site-Specifically Incorporate Methyl-Lysine Analogues into Recombinant Proteins

1 Introduction

2 Protocol to Install MLA into Recombinant Proteins

Chapter Four Methods Applied to the Study of Protein Arginine Methylation

1 Introduction

2 In Vitro Arginine-Methylation Assays

3 Enzyme Sources for In Vitro Methylation Reactions

4 Substrate Sources for In Vitro Methylation Assays

5 Cell-Based Methylation Assays

6 Methyl-Specific Antibodies

7 Using Global-Methylation Inhibitors

8 Using Cell-Based Reporter Systems to Screen for PRMT Inhibitors

9 Summary

Chapter Five Enzymatic Analysis of Tet Proteins: Key Enzymes in the Metabolism of DNA Methylation

1 Introduction

2 Expression and Purification of Tet Proteins

3 Tet Activity Assays

4 Analysis of the Cytosine Derivatives b

Details

No. of pages:
390
Language:
English
Copyright:
© 2012
Published:
Imprint:
Academic Press
eBook ISBN:
9780123919410
Print ISBN:
9780123919403

About the serial-volume-editors

Carl Wu

Affiliations and Expertise

National Cancer Institute, Bethesda, MD

C. Allis

Affiliations and Expertise

The Rockefeller University, New York, NY

Reviews

This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. The first of 2 volumes covering nucleosomes, histones and chromatin, it has chapters on methods applied to the study of protein arginine methylation, high-resolution identification of intra- and interchromosomal DNA interactions by 4C technology, and peptide arrays to interrogate the binding specificity of chromatin-binding proteins.