Glycosyltransferases - Methods and Protocols

Glycosyltransferases (GTs) are essential for the biosynthesis of complex glycoconjugates and are powerful tools to study the functions of complex glycans in health, development and disease. Complex glycoconjugates, such as glycoproteins, proteoglycans and glycolipids, are assembled by GTs which synthesize specific linkages between sugars or sugars and protein. This is in contrast to the non-specific or less specific chemical glycation reactions, transglycosylation and reverse glycosylation reactions. Glycosyltransferases: Methods and Protocols contains a wide range of studies, methods and protocols which form a solid basis for investigations of the role and mechanisms, biology and pathology involving GTs. Written in the successful Methods in Molecular Biology(TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Glycosyltransferases: Methods and Protocols is a vital contribution to glycobiology and glycopathology, and to applications of these enzymes in biotechnology and drug development. It will prove invaluable to students, postdoctoral fellows, and senior scientists carrying on research of GTs that has been intensified over the last years.

List of contents

Glycan Microarray Screening Assay for Glycosyltransferase Specificities.- A Fluorescence-Based Assay for Core 1 3Galactosyltransferase (T-synthase) Activity.- Structural and Biochemical Analysis of a Bacterial Glycosyltransferase.- Study of the Biological Functions of Mucin Type Core 3 O-glycans.- Generation of Anti-Sulfated Glycan Antibodies using Sulfotransferase-deficient Mice.- Fluorescent Microscopy as a Tool to Elucidate Dysfunction and Mislocalization of Golgi Glycosyltransferases in COG Complex Depleted Mammalian Cells.- A Practical Approach to Reconstruct Evolutionary History of Animal Sialyltransferases and Gain Insights into the Sequence-Function Relationships of Golgi-Glycosyltransferases.- Fluorescent Lectin Staining of Drosophila Embryos and Tissues to Detect the Spatial Distribution of Glycans during Development.- Photoaffinity Labeling of Protein O-Mannosyltransferases of the PMT1/PMT2 Subfamily.- Enzymatic Analysis of the Protein O-glycosyltransferase, Rumi, Acting toward Epidermal Growth Factor-like (EGF) Repeats.- Enzymatic Characterization of Recombinant Enzymes of O-GlcNAc Cycling.- Antibodies and Activity Measurements for the Detection of O-GlcNAc Transferase, and Assay of its Substrate, UDP-GlcNAc.- In vitro Glycosylation Assay for Bacterial Oligosaccharyltransferases.- In vitro UDP-sugar:Undecaprenyl-Phosphate Sugar-1-Phosphate Transferase Assay and Product Detection by Thin Layer Chromatography.- In vitro O-Antigen Ligase Assay.- Functional Identification of Bacterial Glucosyltransferase WbdN.- Methods for the Pasteurella Glycosaminoglycan Synthases: Enzymes that Polymerize Hyaluronan, Chondroitin, or Heparosan Chains.- Methods for Measuring Class I Membrane-bound Hyaluronan Synthase Activity.- Creation and Characterization of Glycosyltransferase Mutants of Trypanosoma brucei.- Mannose-6-Phosphate: A Regulator of LLO Destruction.- N-Acetylglucosaminyltransferase (GnT) Assays using Fluorescent Oligosaccharide Acceptor Substrates:GnT-III, IV, V, and IX (GnT-Vb).- A Method for Determination of UDP-GlcNAc: GlcNAcbeta1-6(GlcNAcbeta1-2)Man 1-R [GlcNAc to Man] beta1-4N-acetylglucosaminyltransferase VI Activity.- In vitro Assays of Orphan Glycosyltransferases and their Application to Identify Notch Xylosyltransferases.- In vitro Folding of -1,4Galactosyltransferase and Polypeptide- -N-acetylgalactosaminyltransferase from the Inclusion Bodies.- An Assay for 1,6-Fucosyltransferase (FUT8) Activity Based on the HPLC Separation of a Reaction Product with Fluorescence Detection.- Immunodetection of Glycosyltransferases in Gastrointestinal Tissues.- Glycosyltransferases in Chemo-Enzymatic Synthesis of Oligosaccharides.- Tumor Targeting by a Carbohydrate Ligand-Mimicking Peptide.- Glycoengineering of Human Cell Lines Using Zinc Finger Nuclease Gene Targeting - SimpleCells with Homogeneous GalNAc O-glycosylation Allow Isolation of the O-glycoproteome by One-step Lectin Affinity Chromatography.- Generation of Monoclonal Antibodies to Native Active Human Glycosyltransferases.

Summary

Glycosyltransferases (GTs) are essential for the biosynthesis of complex glycoconjugates and are powerful tools to study the functions of complex glycans in health, development and disease.  Complex glycoconjugates, such as glycoproteins, proteoglycans and glycolipids, are assembled by GTs which synthesize specific linkages between sugars or sugars and protein. This is in contrast to the non-specific or less specific chemical glycation reactions, transglycosylation and  reverse glycosylation reactions. Glycosyltransferases: Methods and Protocols contains a wide range of studies, methods and protocols which form a solid basis for investigations of the role and mechanisms, biology and pathology involving GTs. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.
 
Authoritative and easily accessible, Glycosyltransferases: Methods and Protocols is a vital contribution to glycobiology and glycopathology, and to applications of these enzymes in biotechnology and drug development. It will prove invaluable to students, postdoctoral fellows, and senior scientists carrying on research of GTs that has been intensified over the last years.

Product details

• Assisted by: Ink Brockhausen (Editor), Inka Brockhausen (Editor)
• Publisher: Springer, Berlin
• Languages: English
• Product format: Paperback / Softback
• Released: 01.01.2016
• EAN: 9781493959969
• ISBN: 978-1-4939-5996-9
• No. of pages: 425
• Dimensions: 179 mm x 255 mm x 26 mm
• Weight: 857 g
• Illustrations: XVII, 425 p. 110 illus., 40 illus. in color.
• Series: Methods in Molecular Biology; Methods in Molecular Biology
• Subjects: Natural sciences, medicine, IT, technology > Biology > Biochemistry, biophysics; B, ORGANIC CHEMISTRY, biochemistry, CARBOHYDRATES, Biomedical and Life Sciences, Biochemistry, general, Carbohydrate Chemistry, Analytical Chemistry