Glycobiology / edited by Minoru Fukuda.

Format:

Book

Contributor:

Fukuda, Minoru, 1945-

Series:

Methods in enzymology ; v. 415.

Methods in enzymology ; v. 415

Language:

English

Subjects (All):

Carbohydrates.

Enzymes.

Physical Description:

1 online resource (422 p.)

Place of Publication:

Amsterdam ; Boston : Elsevier/Academic Press, c2006.

Language Note:

English

Summary:

In the past decade, there has been an explosion of progress in understanding the roles of carbohydrates in biological systems. This explosive progress was made with the efforts in determining the roles of carbohydrates in immunology, neurobiology and many other disciplines, examining each unique system and employing new technology. This volume represents the first of three in the Methods in Enzymology series, including Glycomics (vol. 416) and Functional Glycomics (vol. 417), dedicated to disseminating information on methods in determining the biological roles of carbohydrates. These bo

Contents:

Front Cover; Glycobiology; Copyright Page; Table of Contents; Contributors; Preface; Volumes in Series; Section I: N-Glycan Processing; Chapter 1: Non-Radioactive Analysis of Lipid-Linked Oligosaccharide Compositions by Fluorophore-Assisted Carbohydrate Electrophoresis; Overview; Materials; Procedures; Acknowledgments; References; Chapter 2: Identification of N-Glycan-Binding Proteins for E3 Ubiquitin Ligases; Overview; Binding Protein Assay for N-Glycans; In Vitro Ubiquitination Assay for N-linked Glycoproteins with SCFFbx2; Overlay Assay for the Detection of Fbx2 Binding Proteins

Pull-Down Assay for the Interaction Between Fbx2 and N-Linked GlycoproteinsInteraction of Fbx2 with Integrin ß1 in Neuro2a Cells; Acknowledgments; References; Chapter 3: Family 47 alpha-Mannosidases in N-Glycan Processing; Overview; Overview of Class 1 (GH47) Expression and Purification; Generation of Class 1 alpha-Mannosidase Site-Directed Mutants; Enzyme Expression and Purification; Oligosaccharide Isolation for Enzyme Assays and Binding Studies; alpha-Mannosidase Assays and Kinetic Analyses; Glycan Binding Studies by SPR; Summary; Acknowledgment; References

Chapter 4: A Cytoplasmic Peptide: N-GlycanaseIntroduction; In Vitro PNGase Assay; Glycoprotein Substrate for In Vivo PNGase Assay; Technical Considerations; Acknowledgments; References; Section II: Structural Analysis; Chapter 5: Glycomic Profiling of Cells and Tissues by Mass Spectrometry: Fingerprinting and Sequencing Methodologies; Overview; Preparation of Glycans for Mass Spectrometric Analysis; Enzymatic and Chemical Degradations; Derivatization and Purification of Derivatized Glycans; MS Analyses; Automated Analysis of N-Glycan MALDI Fingerprints; References

Chapter 6: Structural Analysis of Sialyl N-Glycan Using Pyridylamination and Chromatography Followed by Multistage Tandem Mass SpectrometryOverview; PA-Oligosaccharide Preparation; 3-DM Using HPLC; Determination of Structure Using Databases and Partial Degradation; Structural Assignment of MSn Spectral Matching; Acknowledgments; References; Chapter 7: Determination of Glycosylation Sites and Disulfide Bond Structures Using LC/ESI-MS/MS Analysis; Overview; Materials; Methods; Identifying Cys-Containing Peptides and N-Linked Glycopeptides; Identifying Disulfide-Bonded Pairs of Peptides

Alkylation of Free Cys Residues (Protocol 1)Peptide: N-Glycosidase (PNGase) F Digestion (Protocol 2); Digestion with Trypsin, Chymotrypsin, or Endoproteinase Glu-C (Protocol 3); Identification of Free Cys-Containing and Disulfide-Bonded Peptides by LC/ESI-MS/MS (Protocol 4); Complete Reduction with DTT and Alkylation with IAM (Protocol 5); Notes; Acknowledgments; References; Chapter 8: Identification of O-GlcNAc Sites on Proteins; Introduction; Immunoblot Detection of O-GlcNAc; Site Mapping by Galactosyltransferase Labeling and Edman Degradation; Autogalactosylation of Galactosyltransferase

Galactosyltransferase Labeling

Notes:

Description based upon print version of record.

Includes bibliographical references and indexes.

ISBN:

9786610751426

9781280751424

1280751428

9780080468990

0080468993

OCLC:

808601202