ADAMTS Proteases Methods and Protocols

This volume provides a variety of methods used to analyze ADAMTS proteases and ADAMTS-like proteins, including their structure, substrate profile, tissue and cell distribution, post-translational modification and biological pathways. Chapters discuss genetic, cell biology, biochemical and proteomics techniques applicable to the analysis of all the members of the ADAMTS superfamily. 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.

 

Cutting-edge and thorough, ADAMTS Proteases: Methods and Protocols is a valuable tool for researchers to study both simple and sophisticated methodologies that are often difficult to locate in primary research literature. 

ADAMTS Proteins: Concepts, Challenges, and Prospects.- Inhibition of ADAMTS1 Expression by Lentiviral CRISPR/Cas9 Gene Editing Technology.- Analyzing the Effects of O-Fucosylation on Secretion of ADAMTS Proteins using Cell-Based Assays.- Challenges and Solutions for Purification of ADAMTS Proteases: An Overview.- Purification of Native or Recombinant ADAMTS2 and Procollagen I Cleavage Assay.- ADAMTS7: Recombinant Protein Expression and Purification.- Purification and Activity Determination of ADAMTS-4 and ADAMTS-5 and Their Domain Deleted Mutants.- Expression and Preparation of Recombinant Reelin and ADAMTS-3 Proteins.- Assay for Reelin-Cleaving Activity of ADAMTS and Detection of Reelin and Its Fragments in the Brain.- Chemical Modification of Proteoglycanases with Biotin.- Analysis of Aggrecanase Activity using Neoepitope Antibodies.- Molecular Cloning, Lentiviral Transduction, and Expression of Recombinant ADAMTSL2 and ADAMTSL4.- Purification of Recombinant ADAMTSL2.- Expression Analysis by RNAscope™ In Situ Hybridization.- Analysis of ADAMTS Effects on Cell Adhesion and Migration.- Cell-Based Interaction Analysis of ADAMTS Proteases and ADAMTS-Like Proteins with Fibrillin Microfibrils.- Alizarin Red and Alcian Blue Preparations to Visualize the Skeleton.- Biophysical Techniques to Analyze Elastic Tissue Extracellular Matrix Proteins Interacting with ADAMTS Proteins.- Quantification of Extracellular Matrix Fiber Systems Related to ADAMTS Proteins.- Visualization of Perineuronal Nets in Central Nervous System Tissue Sections.- Visualization and Quantification of Pericellular Matrix.- Quantitative Mass Spectrometry-Based Secretome Analysis as a Tool to Investigate Metalloprotease and TIMP Activity.- Preparation and Application of a Decellularized Extracellular Matrix for Identification of ADAMTS Substrates.- Mapping the N-Terminome in Tissue Biopsies by PCT-TAILS.