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This volume is a wide-ranging tool for studying protein-carbohydrate interactions that extend from traditional biochemical methods to state-of-the-art techniques. This book focuses on four different research themes: Part I describes methods for screening and quantifying CAZyme activity; Part II contains methods for investigating the interactions between proteins and carbohydrate ligands; Part III discusses methods for the visualization of carbohydrates and protein-carbohydrate complexes; and Part IV focuses on structural and "omic" approaches for studying systems of CAZymes. 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, Protein-Carbohydrate Interactions: Methods and Protocols is a valuable resource to the glycomics research community. In this continuously advancing field, the methods in this book highlight the biology of glycomics, thus driving biotechnological innovation and solutions for human health and sustainable resources within the emerging green community.
A Low-Volume, Parallel Copper-Bicinchoninic Acid (BCA) Assay for Glycoside Hydrolases.- Quantitative Kinetic Characterization of Glycoside Hydrolases using High-Performance Anion-Exchange Chromatography (HPAEC).- Measuring Enzyme Kinetics of Glycoside Hydrolases using the 3,5-Dinitrosalicylic Acid Assay.- An Improved Kinetic Assay for the Characterization of Metal-Dependant Pectate Lyases.- Colorimetric Detection of Acetyl Xylan Esterase Activities.- Methods for Determining Glycosyltransferase Kinetics.- Analyzing Activities of Lytic Polysaccharide Monooxygenases by Liquid Chromatography and Mass Spectrometry.- Carbohydrate Depolymerization by Intricate Cellulosomal Systems.- Affinity Electrophoresis for Analysis of Catalytic Module-Carbohydrate Interactions.- Quantifying CBM Carbohydrate Interactions using Microscale Thermophoresis.- Characterization of Protein-Carbohydrate Interactions by NMR Spectroscopy.- Measuring the Biomechanical Loosening Action of Bacterial Expansins on Paper and Plant Cell Walls.- Bioinspired Assemblies of Plant Cell Walls for Measuring Protein-Carbohydrate Interactions by FRAP.- CBMs as Probes to Explore Plant Cell Wall Heterogeneity using Immunocytochemistry.- Determining the Localization of Carbohydrate Active Enzymes within Subcellular Compartments of Gram-Negative Bacteria.- Analysis of Complex Carbohydrate Composition in Plant Cell Wall using Fourier Transformed Mid-Infrared Spectroscopy (FT-IR).- Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis.- A Rapid Procedure for the Purification of 8-Aminopyrene Trisulfonate (APTS)-Labelled Glycans for Capillary Electrophoresis (CE)-Based Enzyme Assays.- Probing the Complex Architecture of Multimodular Carbohydrate-Active Enzymes using a Combination of Small Angle X-Ray Scattering and X-Ray Crystallography.- Metagenomics and CAZyme Discovery.- Identification of Genes Involved in the Degradation of Lignocellulose using Comparative Transcriptomics.- Isolation and Preparation of Extracellular Proteins from Lignocellulose Degrading Fungi for Comparative Proteomic Studies using Mass Spectrometry.
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