Glycoprotein Methods and Protocols: The Mucins

ISBN-13: 9781617371493 / Angielski / Miękka / 2010 / 506 str.

Anthony P. Corfield

Opis

The mucins (mucus glycoproteins) have long been a complex corner of glycoprotein biology. While dramatic advances in the separation, structural an- ysis, biosynthesis, and degradation have marked the progress in general glycop- tein understanding, the mucins have lagged behind. The reasons for this lack of progress have always been clear and are only now being resolved. The mucins are very large molecules; they are difficult to separate from other molecules present in mucosal secretions or membranes; they are often degraded owing to natural protective functions or to isolation methodology and their peptide and oligos- charide structures are varied and complex. Understanding these molecules has demanded progress in several major areas. Isolation techniques that protect the intact mucins and allow dissociation from other adsorbed but discrete molecules needed to be developed and accepted by all researchers in the field. Improved methods for the study of very large molecules with regard to their aggregation and polymerization were also needed. Structural analysis of the peptide domains and the multitude of oligosaccharide chains was required for smaller sample sizes, for multiple samples, and in shorter time. In view of these problems it is perhaps not surprising that the mucins have remained a dilemma, of obvious biological importance and interest, but very difficult to analyze.

Recenzje

"There is sufficient detail to follow easily and unlike many research papers, the minute details which make for success of failure, all appear to included." -Microbiology Today

Spis treści

Part I. Purification of Mucin. Isolation of Large Gel-Forming Mucins, Julia R. Davies and Ingemar Carlstedt. Preparation of Membrane Mucin, Kermit L. Carraway. Part II. Detection and Quantitation of Mucin. Histologically Based Methods for Detection of Mucin, Michael D. Walsh and Jeremy R. Jass. Detection and Quantitation of Mucins Using Chemical, Lectin, and Antibody Methods, Michael A. McGuckin and David J. Thornton. The Gastrointestinal Adherent Mucous Gel Barrier, Adrian Allen and Jeffrey P. Pearson. Quantitation of Biosynthesis and Secretion of Mucin Using Metabolic Labeling, Jan Dekker, B. Jan-Willem Van Klinken, Hans A. Büller, and Alexandra W. C. Einerhand. Part III. Separation, Identification, and Physical Characterization of Mucins. Separation and Identification of Mucins and Their Glycoforms, David J. Thornton, Nagma Khan, and John K. Sheehan. Heterogeneity and Size Distribution of Gel-Forming Mucins, John K. Sheehan and David J. Thornton. Part IV. Rheology of Mucin. Rheology of Mucin, Jeffrey P. Pearson, Adrian Allen, and David A. Hutton. Part V. Mucin Peptide Analysis. Amino Acid Analysis of Mucins, Jun X. Yan and Nicolle H. Packer. Identification of Glycosylation Sites in Mucin Peptides by Edman Degradation, Natasha E. Zachara and Andrew A. Gooley. Synthetic Peptides for the Analysis and Preparation of Antimucin Antibodies, Andrea Murray, Deirdre A. O'Sullivan, and Michael R. Price. Mucin Domains to Explore Disulfide-Dependent Dimer Formation, Sherilyn L. Bell and Janet F. Forstner. Part VI. Carbohydrate Structural Analysis of Mucins. Monosaccharide Composition of Mucins, Jean-Claude Michalski and Calliope Capon. O-Linked Oligosaccharide Chain Release and Fractionation, Elizabeth F. Hounsell. Structural Analysis of Mucin-Type O-Linked Oligosaccharides, André Klein, Gérard Strecker, Geneviève Lamblin, and Philippe Roussel. Measurement of Sulfate in Mucins, Mathew J. Harrison and Nicolle H. Packer. Part VII. Mucin Biosynthesis. Biosynthesis of MucinCell and Organ Culture Methods for Biosynthetic Study, Anthony P. Corfield, Neil Myerscough, Alexandra W. C. Einerhand, B. Jan-Willem Van Klinken, Jan Dekker, and Christos Paraskeva. Metabolic Labeling Methods for the Preparation and Biosynthetic Study of Mucin, Anthony P. Corfield, Neil Myerscough, B. Jan-Willem Van Klinken, Alexandra W. C. Einerhand, and Jan Dekker. Identification of Mucins Using Metabolic Labeling, Immunoprecipitation, and Gel Electrophoresis, B. Jan-Willem Van Klinken, Hans A. Büller, Alexandra W. C. Einerhand, and Jan Dekker. Mucin Precursors: Identification and Analysis of Their Intracellular Processing, Alexandra W. C. Einerhand, B. Jan-Willem Van Klinken, Hans A. Büller, and Jan Dekker. Inhibition of Mucin Glycosylation, Guillemette Huet, Philippe Delannoy, Thécla Lesuffleur, Sylviane Hennebicq, and Pierre Degand. O-Linked Chain Glycosyltransferases, Inka Brockhausen. Part VIII. Mucin Gene Detection. Mucin cDNA Cloning, Jean-Pierre Aubert and Nicole Porchet. Northern Blot Analysis of Large mRNAs, Nicole Porchet and Jean-Pierre Aubert. Southern Blot Analysis of Large DNA Fragments, Nicole Porchet and Jean-Pierre Aubert. In Situ Hybridization Techniques for Localizing Mucin mRNA, Ilene K. Gipson. Detection of Mucin Gene Polymorphism, Lynne E. Vinall, Wendy S. Pratt, and Dallas M. Swallow. Part IX. Preparation of Antimucin Antibodies. Polyclonal and Monoclonal Techniques, Francisco X. Real, Carme de Bolós, and Egbert Oosterwijk. Monoclonal Antibodies to Mucin VNTR Peptides, Pei Xiang Xing, Vasso Apostolopoulos, Jim Karkaloutsos, and Ian F. C. McKenzie. Part X Enzymatic Degradation of Mucins. Mucinase Activity, Roger M. Stark, Rebecca Wiggins, Elizabeth Walley, Sally J. Hicks, Gulnaz A. Gill, Stephen D. Carrington, and Anthony P. Corfield. Proteinase Activity, David A. Hutton, Adrian Allen, and Jeffrey P. Pearson. Glycosidase Activity, Anthony P. Corfield and Neil Myerscough. Assays for Bacterial Mucin-Desulfating Sulfatases, Anthony M.

Blurb

Anthony P. Corfield and a group of hands-on expermentalists bring together in Glycoprotein Methods and Protocols: The Mucins a state-of-the-art collection of reliable and tested methods for the study of all aspects of mucins. These powerful methods range from the preparation of mucins to their detection and quantitation with molecular biological and biochemical reagents. Also included are methods for the assessment of mucin peptide and carbohydrate, of mucin biosynthesis and degradation, and of mucin bacteriology and cell biology. The techniques take advantage of the latest improvements in sensitivity and specificity of detection, and in the preparation of new reagents for specific biochemical detection. Glycoprotein Methods and Protocols: The Mucins offers today's researchers a complete range of readily reproducible analytical techniques for studying mucins. Its powerful techniques constitute a launching pad for those entering the field for the first time and a state-of-the-art collection for those already active in mucin research.