Abstract

Techniques in Glycobiology

(Townsend, R. R., and Hotchkiss, A. T., Jr. (eds.) Marcel Dekker, New York-Basel-Hong Kong, 1997, 767 p.)

The book is a practical manual on application of modern techniques to studies of complex carbohydrates and glycoconjugates. The book consists of 6 parts which are divided to separate chapters written by well-known specialists from different countries.

Part I is devoted to mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. Chapter 1 (A. E. Manzi and P. A. Keifer) describes a new NMR spectroscopy procedure which allows analysis of nanogram quantities of oligosaccharides. Using so-called nano-NMR probes, the authors could identify a new oligosaccharide sequence in a glycosaminoglycan fraction (25 µg) isolated from human melanoma cells. Chapter 2 (B. W. Gibson et al.) presents new approaches to structure elucidation of bacterial polysaccharides using matrix-assisted laser desorption/ionization time-of-flight MS. Chapter 3 (D. I. Papac et al.) illustrates application of this method to determination of molecular mass of N-linked oligosaccharides from recombinant plasminogen activator. Oligosaccharides were separated by high-performance anion-exchange chromatography (HPAEC) at high pH using Dionex columns. Chapters 4 (U. Rapp) and 5 (P. A. Haynes et al.) give an account of structures of glycopeptides and unusual phosphorylated oligosaccharides from Trypanosoma cruzi. Chapters 6 (Y. Ohashi) and 7 (C. Delmas) describe applications of MS and NMR spectroscopy, respectively, to studies of carbohydrate--protein interactions and structure of bacterial polysaccharides.

Part II which includes 7 chapters (8-14), centers on new physical methods of ganglioside analysis. Chapter 8 (T. Taki) is concerned with microscale analysis of glycosphingolipids by thin-layer chromatography (TLC) followed by transfer of separated glycosphingolipids to a special membrane. High-sensitivity identification of glycosphingolipids was achieved using monoclonal antibodies. Chapter 9 (S. Sonnino et al.) deals with methods of separation of gangliosides using high-performance liquid chromatography (HPLC). Chapter 10 (M. H. Ravindranath et al.) describes a new highly sensitive method of quantitative determination of gangliosides and cell surface carbohydrate antigens using monoclonal antibodies and immunosorbents. Chapter 11 (L. Cantu et al.) gives examples of modification of cell membrane glycolipids, which may be controlled using various physical methods, such as laser and cold neutron scattering. Chapter 12 (T. M. McIntire and D. A. Brant) contains data on molecular structure of plant and bacterial polysaccharides obtained by noncontact atomic-force microscopy. Two new biosensor methods for analysis of carbohydrate--protein interactions are discussed in chapters 13 (Y. Shinohara et al.) and 14 (K. Dill).

Part III covers various techniques. Chapter 15 (T. A. Gerken et al.) gives notions on O-glycosylation effects on conformation of polypeptides containing O-linked oligosaccharide chains. Computer algorithms for predicting O-glycosylation sites in glycoproteins are considered in chapter 16 (E. P. Elhammer et al.). Chapter 17 (K. Nehrke et al.) which is logically connected with chapter 16, presents new approaches to studying O-glycosylation in vivo. Methods of identifying sites of O-glycosylation, such as MS, gas chromatography, and their combination, are examined in detail in chapters 18 (A. Pisano et al.), 19 (K. F. Medzihradszky), and 20 (K. A. Thomsson).

Techniques of labeling oligosaccharides with chromophores and fluorophores are treated in part IV of the book. Chapters 21 (K. R. Anumula), 22 (M. R. Hardy), and 23 (A. Guttman) center on various procedures of modification of oligosaccharide chains of glycoproteins for fluorescent labeling. The labeling increases significantly the sensitivity of detection of carbohydrate components after separation using chromatography and electrophoresis, including capillary electrophoresis. Chapter 24 (W. C. Kett et al.) describes a procedure for determination of glycopyrazole derivatives of sugars and its application to structural analysis of glycoconjugates.

Part V is devoted to methods of separation and structure elucidation of oligosaccharides. Data on the specificity of various glycosidases useful for structural analysis of glycoconjugates are dealt with in chapter 25 (S. Wong-Madden et al.). TLC of coumarin-labeled products of enzymatic hydrolysis allows fast and highly sensitive screening of the specificity of glycosidases from various sources. Chapter 26 (M. Stefansson and M. Novotny) summarizes methods of resolution of oligosaccharides by capillary electrophoresis. Chapter 27 (E. Higgins and R. Bernasconi) compares various methods of separation of oligosaccharides, such as HPAEC at high pH, electrophoresis of fluorogen-labeled carbohydrates, and HPLC. New conditions of sample preparation for efficient separation of oligosaccharides using HPAEC at high pH are discussed in chapter 28 (P. H. Lipniunas et al.).

Chapter 29 (M. J. Rao et al.) covers various chromatographic approaches to monosaccharide analysis of recombinant glycoproteins. Chapters 30 (K. Kopp et al.) and 31 (R. Nuck and M. Gohlke) describe procedures of 2-aminobenzamide labeling of desialylated oligosaccharides from recombinant glycoproteins. Such labeling allows determination of subnanomolar quantities of N-glycans using MS and laser MS. Chapter 32 (R. B. Trimble) deals with methods of isolation and characterization of N-linked oligosaccharides from yeast glycoproteins.

Part VI of the book is dedicated to plant glycobiology. Chapter 33 (M. G. O'Shea and M. K. Morell) describes a new electrophoresis procedure for resolution of carbohydrates using a DNA sequencer. Recent progress in starch structure analysis is discussed in chapter 34 (K.-S. Wong and J. Jane). Special attention is given to HPAEC with pulsed amperometric detection. Methods of structural analysis of seaweed galactans are the subject of chapter 35 (C. A. Stortz et al.). Chapter 36 (N. C. Carpita and M. C. McCann) is concerned with new methods to study plant polyuronic acids and their esters. Authors consider some of these methods as useful for analysis of specific organization of pectins and dynamics of their development in a separate cell wall. Chapter 37 (M. R. Gretz et al.) which completes the book, examines some methods for determination of the structure and location of cellulose and carrageenans in red algal extracellular matrix.

The book as a whole is a highly valuable manual covering most modern approaches to study glycoconjugates. Being written by a group of more than 100 worldwide-known experts, it will rightly become a handbook not only for glycobiologists, but also for specialists in the fields of bioorganic chemistry, biochemistry, molecular biology, and pharmacology. It will become helpful for teachers and students of universities and biotechnological institutes as well.

Bibliography to each chapter (there are more than 1300 references in the total) and a subject index supplement the book and enable fast orientation within the voluminous data presented.

G. Ya. Wiederschain