MMPs and TIMPs-An Overview of the Field -- MMPs and TIMPs -- Strategies for Cloning New MMPs and TIMPs -- Structural Studies on MMPs andTIMPs Wolfram Bode and Klaus Maskos -- Matrix MetaIIoproteinase Substrate Binding Domains, Modules and Exosites -- The Matrix Metalloproteinase (MMP) and Tissue Inhibitor of Metalloproteinase (TIMP) Genes -- Models for Gain-of-Function and Loss-of-Function of MMPs -- Expression and Purification of MMPs and TIMPs -- Expression of MMPs andTIMPs in Mammalian Cells -- Expression of Recombinant Matrix Metalloproteinases in Escherichia coli -- Expression of Human Collagenase I (MMP-1) andTIMP-1 in a Baculovirus-Based Expression System -- Expression of Recombinant Matrix Metalloproteinases in Yeast -- Expression of Recombinant Membrane-Type MMPs -- Refolding of TIMP-2 from Escherichia coli Inclusion Bodies -- Expression and Refolding of Full-Length HumanTIMP-1 -- Purification of MMPs and TIMPs -- Detection of MMPs and TIMPs -- Monitoring MMP andTIMP mRNA Expression by RT-PCR -- Measuring Transcription of Metalloproteinase Genes -- In Situ Hybridization for Metalloproteinases and Their Inhibitors -- Use of EIA to Measure MMPs and TIMPs -- Immunohistochemistry of MMPs and TIMPs -- Detecting Polymorphisms in MMP Genes -- Assay of MMP and TIMP Activities -- Methods for Studying Activation of Matrix Metalloproteinases -- Assay of Matrix Metalloproteinases Against Matrix Substrates -- Zymography and Reverse Zymography for Detecting MMPs, andTIMPs -- In Situ Zymography -- Detection of Focal Proteolysis Using Texas-Red-Gelatin -- Antibodies to MMP-Cleaved Aggrecan -- Cartilage Proteoglycan Release Assay -- Immunoassay for Collagenase-Mediated Cleavage of Types I and II Collagens -- Collagen Degradation Assays -- Invasion Assays and Matrix Metalloproteinases -- Using Fluorogenic Peptide Substrates to Assay Matrix Metalloproteinases -- Kinetic Analysis of the Inhibition of Matrix Metalloproteinases by Tissue Inhibitor of Metalloproteinases (TIMP) -- Assaying Growth Factor Activity of Tissue Inhibitors of Metalloproteinases.

From the simple discovery in 1962 that resorbing tadpole tail expressed an enzyme (MMP) that could degrade collagen gels, matrix metalloproteinase (MMP) research has advanced to discover more than twenty distinct vertebrate MMPs and four specific inhibitors (TIMPS), a veritable family of enzymes involved in many physiological and pathological processes. In Matrix Metalloproteinase Protocols, leading experts detail proven laboratory techniques for the study of MMPs. The methods include those for the expression and purification of MMPs and TIMPs, for the detection of MMPs and TIMPs at both the protein and mRNA levels, and for the assay of MMP and TIMP activities in a wide variety of circumstances. Each method includes step-by-step instructions and notes on variant applications and pitfalls to avoid. A selective overview of the MMP arena spells out where the field has been, where it is, and where it is going. Comprehensive and highly practical, Matrix Metalloproteinase Protocols brings together the long and hard-earned experience of master experimentalists that will allow not only novices to get up to speed quickly, but also add to the repertoire of successful techniques in expert laboratories.