Titlebook: Matrix Metalloproteinase Protocols; Ian M. Clark Book 2010Latest edition Humana Press 2010 Escherichia coli.Microarray.Nucleotide.PCR.Prot

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Methods for Studying Activation of Matrix Metalloproteinasesyme systems, including the matrix metalloproteinases (MMPs), serine proteinases, and cysteine proteinases. The majority of the soluble MMPs are synthesized as proenzymes which require extracellular activation in order to gain proteolytic activity and the analysis of their activation mechanism is a p

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Assay of Matrix Metalloproteinases Against Matrix Substrateso be measured. The protocols described include the preparation of radiolabeled substrates, methods for the separation of degraded product from undegraded substrate, and methods for the activation of MMPs. The advantages and disadvantages of these methods are discussed in relation to immunoassays tha

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Metalloproteases and the Degradomeree families of metalloproteases: ADAMs (a disintegrin and metalloproteinase), ADAMTSs (ADAMs with thrombospondin domains), and MMPs (matrix metalloproteinases) which have a growing relevance in a number of human pathologies including cancer, arthritis, neurodegenerative disorders, and cardiovascular diseases.

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Expression of Recombinant ADAMTS in Insect Cellsologies related to increased turnover of the extracellular matrix, particularly in tissues where the concentration of proteoglycans is high, such as cartilage and the central nervous system. We have expressed three of these enzymes, ADAMTS-1, -4 and -5, in insect cells using plasmid-based systems.

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Refolding of TIMP-2 from , Inclusion Bodiesintracellular expression and efficient isolation of the recombinant product without the use of fusion tags or partners. Protein purity after ion exchange and gel filtration chromatography was judged to be greater than 95% with yields of 15?mg/L from LB medium and 10?mg/L from minimal medium.

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Methods for Studying Activation of Matrix Metalloproteinasesrerequisite for understanding MMP-mediated proteolysis..The emphasis of this chapter is the provision of the experimental tools to study MMP activation in vitro and in cellular model systems. Hence, we use the activation of procollagenase-3 (proMMP-13) and progelatinase A (proMMP-2) as examples of the methods used.

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Analysis of the Degradome with the CLIP-CHIP? Microarraych parts of the sequence of known or predicted protease and inhibitor mRNAs in both species and printed on a glass-matrix surface, the CLIP-CHIP? microarray can be used to analyze differentially expressed protease and inhibitor gene products and give expression profiles for any analyzed sample.