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This unique book covers the latest developments in coupling and decoupling of biomolecules containing functionalized carbohydrate components, being one of the first collections in this important area of applied medicinal chemistry. Connecting molecules, often referred as bio-conjugation, has become one of the most often performed procedures in modern medicinal chemistry. Sometimes, when the connected molecules are not useful anymore, they must be disconnected. The molecules that must be connected (coupled) may belong to both small and large molecules and include such constructs as glycoproteins, glycopeptides and glycans. In this work, more than 15 experts address a comprehensive range of potential and current uses of in vitro and in vivo bio-conjugation methodologies, leading to a variety of glycoconjugates. The analytical aspects of bio-conjugation are also here discussed. Medicinal and organic chemists from graduate level onwards will understand the appeal of this important book.
By guiding the reader toward answering basic questions on the structural analysis of glycoconjugates using conventional techniques, this book studies methods such as chemical cleavage, molar composition, use of enzymes, use of lectins and typical fractionation procedures such as HPLC, GLC, TLC, and other chromatographic systems. More sophisticated techniques such as NMR or MS are described to give the reader the essential information needed to adapt procedures to his/her requirements. This book will serve as a practical manual designed for scientists not familiar with glycoconjugate methodologies, but whose research work requires contact with analytical problems or metabolic investigations in glycobiology. It is the result of twenty years of experience of summer schools held in a laboratory environment and will acquaint the reader with widely used techniques in the area rather than cataloguing an exhaustive list of methods. This will help any biochemist to develop an experiment with ease.
By guiding the reader toward answering basic questions on the structural analysis of glycoconjugates using conventional techniques, this book studies methods such as chemical cleavage, molar composition, use of enzymes, use of lectins and typical fractionation procedures such as HPLC, GLC, TLC, and other chromatographic systems. More sophisticated techniques such as NMR or MS are described to give the reader the essential information needed to adapt procedures to his/her requirements. This book will serve as a practical manual designed for scientists not familiar with glycoconjugate methodologies, but whose research work requires contact with analytical problems or metabolic investigations in glycobiology. It is the result of twenty years of experience of summer schools held in a laboratory environment and will acquaint the reader with widely used techniques in the area rather than cataloguing an exhaustive list of methods. This will help any biochemist to develop an experiment with ease.
The fields of glycochemistry and glycoscience are rich and varied and where much can be learned from Nature. As Nature is not always able to produce carbohydrates in quantities useful for not only in research but also as therapeutic agents, new ways need to be found to optimize the yield. This book presents an overview of the latest developments in the field of carbohydrates, ranging from de-novo approaches via cyclodextrin chemistry to the synthesis of such highly complex glycoconjugates as glycosphingolipids and GPI anchors. The main emphasis remains on the synthetic aspects making the book an excellent source of information for those already involved in carbohydrate chemistry, as well as for those organic chemists who are beginners in this field. Equally of interest to synthetic chemists, as well as medicinal chemists and biochemists.
This volume and its companion, Volume 339, supplement Volumes 176, 177, 239, and 261. Chapters are written with a "hands-on" perspective. That is, practical applications with critical evaluations of methodologies and experimental considerations needed to design, execute, and interpret NMR experiments pertinent to biological molecules.
Sugar chains (glycans) are often attached to proteins and lipids and have multiple roles in the organization and function of all organisms. "Essentials of Glycobiology" describes their biogenesis and function and offers a useful gateway to the understanding of glycans.
Glycoconjugates Composition: Structure, and Function provides an excellent overview of the composition, biosynthesis, function and structure of the carbohydrate chains of glycoconjugates from higher organisms. It is recommended as a core reference text, providing excellent coverage of the glycoconjugate field.
18. 2 Principle of FACE/Gel Retardation Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 349 18. 3 Labelling of Oligosaccharides with ANTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 350 18. 4 Screening of Carbohydrate Ligands for Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . 352 18. 5 Measurement of Binding Constant for the Interaction Between Protein and ANTS-Labelled Carbohydrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 355 18. 6 Measurement of Binding Constant for the Interaction Between Protein and Native Carbohydrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360 ~ The Application of Capillary Affinity Electrophoresis to the Analysis _ of Carbohydrate-Protein Interactions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 19. 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 19. 2 Principle of CAE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363 19. 3 Determination of Association Constants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364 19. 4 Technical Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 General considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 19. 5 Limitations of the Technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 370 19. 6 Application of CAE to the Analysis of Carbohydrate-Protein Interactions . . . . . . 371 19. 7 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 375 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 377 20. 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 379 Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 380 20. 2 Technical Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 381 20. 3 Sample Detection and Sample Recovery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Autoradiography and staining . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Sample detection by blotting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Semipreparative ACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 390 20. 4 Analysis of Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 391 Measuring sample mobilities - calculating a retardation coefficient . . . . . . . . . . . . 391 Graphical analysis of data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 392 Interpreting ACE patterns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393 Reverse ACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395 20. 5 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 397 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 Subject Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399 XII List of Contributors Nebojsa Avdalovic John T. Gallagher Dionex Corporation Cancer Research Campaign Department of Medical Oncology 445 Lakeside Drive University of Manchester Sunnyvale, CA 94086 Christie CRC Research Centre Klaus Biemann Wilmslow Road Department of Chemistry Manchester M20 4BX Massachusetts Institute of Technology UK Cambridge, MA 02139-4307 USA Geoffrey R.
Glycoconjugate Research, Volume I contains the proceedings of the Fourth International Symposium on Glycoconjugates, held in Woods Hole, Massachusetts, in September 1977. Contributors focus on the state of knowledge in the field of glycoconjugates ranging from polysaccharides and glycoproteins to glycolipids, proteoglycans, and all their varieties in plants, animals, and microorganisms. This text is organized into three sections and consists of 87 chapters. After an introductory chapter where the use of glycosidases for the structural analysis of complex carbohydrates is described, the discussion turns to other methods used for elucidating the structure of complex carbohydrates. The chapters that follow focus on the primary structure and conformation of glycans that are N-glycosically linked to peptide chains; isolation of monofuco-disialoganglioside from human and bovine brains; and partial chemical characterization of activated peptides isolated from a single cell suspension of rat colonic secretory cells. The last two sections explore cartilage proteoglycans and the structure-function relationships of glycoconjugates, touching on topics such as link proteins; the biochemical aspects of malignancy in human chondrosarcomas; defensive response of plants to a microbial oligosaccharide with a pheromone-like activity; and the interaction of saccharides with ricin. This book is intended for students and practitioners of chemistry and biochemistry; biology and microbiology; and physiology.