Key Features

This volume in the Methods in Enzymology series comprehensively covers the topic of serpins

With an international board of authors, this volume covers subjects such as Crystallography of serpins and serpin complexes, Serpins as hormone transporters, and Production of serpins using cell free systems


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

Table of Contents

Series Page



Methods in Enzymology

Intracellular Production of Recombinant Serpins in Yeast

1. Introduction

2. Selection of Strain and Expression Plasmid

3. Growth of Yeast

4. Transformation of Yeast

5. Screening Transformants

6. Large-Scale Growth and Induction

7. Lysis

8. Purification

9. Assessing Serpin Activity and Removing Inactive Forms

10. Production of Polymerogenic Serpins

Production of Recombinant Serpins in Escherichia coli

1. Introduction

2. Experimental Procedures for the Production of Serpins in E. coli

3. Some Common Variations Used for Expression of Other Serpins

4. Preparation of AT from Inclusion Bodies

5. Preparation of Soluble AT

Isolation and Characterization of the Nuclear Serpin MENT

1. Introduction

2. Purification of the MENT Protein

3. Protease Inhibition/Serpin Activity

4. MENT Interaction with DNA and Chromatin In Vitro

5. Analysis of MENT Association with Native Chromatin In Situ


Solving Serpin Crystal Structures

1. Introduction

2. Protein Production and Purification

3. Modifications to Aid Crystallization

4. Crystallization

5. Experimental Phasing

6. Molecular Replacement

7. Phase Improvement by Density Modification

8. Refinement and Validation

Crystallography of Serpins and Serpin Complexes

1. Introduction

2. First Glimpses of Serpin Structures

3. The Serpin–Enzyme Complex

4. The Michaelis Complex

5. Conformational Control of Serpins—Antithrombin and Heparin

6. Nonconventional Serpin Complexes

7. Conformational Change and the Formation of the Latent Conformation

8. Abnormal Conformatio


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© 2011
Academic Press
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