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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.
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
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