Mehr lesen
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.
Inhaltsverzeichnis
Expression and Purification of Matrix Metalloproteinases in Escherichia coli.- Expression and Purification of a Matrix Metalloprotease Transmembrane Domain in Escherichia coli.- Heterologous Expression of the Astacin Protease Meprin b in Pichia pastoris.- Structural Studies of Matrix Metalloproteinase by X-Ray Diffraction.- Mapping Lipid Bilayer Recognition Sites of Metalloproteinases and other Prospective Peripheral Membrane Proteins.- Using Small Angle X-ray Scattering (SAXS) to Characterise the Solution Conformation and Flexibility of Matrix Metalloproteinases (MMPs).- Molecular Dynamics Studies of Matrix Metalloproteases.- Determining the Substrate Specificity of Matrix Metalloproteases using Fluorogenic Peptide Substrates.- Time-resolved Analysis of Matrix Metalloproteinase Substrates in Complex Samples.- Identification of Protease Cleavage Sites by Charge-Based Enrichment of Protein N-termini.- Mapping theSubstrate Recognition Landscapes of Metalloproteases using Comprehensive Mutagenesis.- Detection of Matrix Metalloproteinases by Zymography.- Imaging Matrix Metalloproteases in Spontaneous Colon Tumors: Validation by Correlation with Histopathology.- Virtual High-throughput Screening for Matrix Metalloproteinase Inhibitors.- Computational Approaches to Matrix Metalloprotease Drug Design.- A Simple, Adaptable Blood-Brain Barrier Cell Model for Screening Matrix Metalloproteinase Inhibitor Functionality.- Matrix Metalloproteases as Biomarkers of Disease.
Zusammenfassung
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.
