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