Main description:

Since the identification of the first matrix metalloprotease (insterstitial collagenase or MMP-1) more than 20 closely related and evolutionarily conserved vertebrate MMPs have been discovered. Chapters in this volume detail methods and protocols on recent advances in the expression and purification of MMPs, biophysical methods such as X-ray crystallography, NMR spectroscopy and small angle X-ray scattering, computational and experimental methods for determining MMP specificity, methods for detecting MMP activity in vitro and in vivo, and methods for the development and characterization of MMP-based inhibitors as potential therapeutics for the treatment of various diseases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Matrix Metalloproteases: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

PART I Expression and Purification of Matrix Metalloproteases

1. Expression and Purification of Matrix Metalloproteinases in Escherichia coli

Krishna K. Singh, Ruchi Jain, Harini Ramanan, and Deepak K. Saini

2. Expression and Purification of a Matrix Metalloprotease Transmembrane Domain in Escherichia coli

Charles A. Galea

3. Heterologous Expression of the Astacin Protease Meprin b in Pichia pastoris

Dagmar Schlenzig and Stephan Schilling

PART II Structural Characterization of Matrix Metalloproteases

4. Structural Studies of Matrix Metalloproteinase by X-Ray Diffraction

Elena Decaneto, Wolfgang Lubitz, and Hideaki Ogata

5. Mapping Lipid Bilayer Recognition Sites of Metalloproteinases and other Prospective Peripheral Membrane Proteins

Tara C. Marcink, Rama K. Koppisetti, Yan G. Fulcher, and Steven R. Van Doren

6. Using Small Angle X-ray Scattering (SAXS) to Characterise the Solution Conformation and Flexibility of Matrix Metalloproteinases (MMPs)

Louise E. Butt, Robert A. Holland, Nikul S. Khunti, Debra L. Quinn, and Andrew R. Pickford

PART III Computational Simulations of Matrix Metalloproteases

7. Molecular Dynamics Studies of Matrix Metalloproteases

Natalia Diaz and Dimas Suarez

PART IV Determining Matrix Metalloprotease Substrate Specificity

8. Determining the Substrate Specificity of Matrix Metalloproteases using Fluorogenic Peptide Substrates

Maciej J. Stawikowski, Anna M. Knapinska, and Gregg B. Fields

9. Time-resolved Analysis of Matrix Metalloproteinase Substrates in Complex Samples

Pascal Schlage, Fabian E. Egli, and Ulrich auf dem Keller

10. Identification of Protease Cleavage Sites by Charge-Based Enrichment of Protein N-termini

Zon W. Lai and Oliver Schilling

11. Mapping the Substrate Recognition Landscapes of Metalloproteases using Comprehensive Mutagenesis

Colin A. Kretz

PART V Detection of Matrix Metalloproteases

12. Detection of Matrix Metalloproteinases by Zymography

Rajeev B. Tajhya, Rutvik S. Patel, and Christine Beeton

13. Imaging Matrix Metalloproteases in Spontaneous Colon Tumors: Validation by Correlation with Histopathology<

Harvey Hensley, Harry S. Cooper, Wen-Chi L. Chang, and Margie L. Clapper

PART VI Matrix Metalloprotease Inhibitors 14. Virtual High-throughput Screening for Matrix Metalloproteinase Inhibitors Jun Yong Choi, and Rita Fuerst

15. Computational Approaches to Matrix Metalloprotease Drug Design Tanya Singh, B. Jayaram, and Olayiwola Adedotun Adekoya

16. A Simple, Adaptable Blood-Brain Barrier Cell Model for Screening Matrix Metalloproteinase Inhibitor Functionality

Jennifer S. Myers, Joan Hare, and Qing-Xiang Amy Sang

PART VII Matrix Metalloproteases as Biomarkers

17. Matrix Metalloproteases as Biomarkers of Disease

Fernando Luiz Affonso Fonseca, Beatriz da Costa Aguiar Alves, Ligia Ajaime Azzalis, and Thais Moura Gascon Belardo