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[9] 1H nuclear magnetic resonance spectroscopy of carbohydrate chains of glycoproteins

1994; Academic Press; Linguagem: Inglês

10.1016/0076-6879(94)30011-9

1557-7988

H. van Halbeek,

Enzyme Structure and Function

Nuclear magnetic resonance (NMR) spectroscopy provides a powerful, nondestructive means to characterize glycoprotein carbohydrates structurally, and has become an integral part of the current methodology of protein glycosylation site mapping. This chapter discusses 1H NMR spectroscopy as a method for the characterization of the primary structure of N-type and O-type oligosaccharide chains of glycoproteins. The practical aspects of the method, including sample preparation, recording a spectrum (data acquisition), and data processing are emphasized. The usefulness of the structural reporter group concept for interpretation of the 1H NMR spectrum of a carbohydrate in terms of its primary structure is outlined. The chapter illustrates the application of 1H NMR spectroscopy for the structural elucidation of the carbohydrate chains of human neutrophil elastase (HNE) and of recombinant human erythropoietin (rEPO) expressed in Chinese hamster ovary (CHO) cells. The most important advantage of NMR spectroscopy over other techniques used for structural analysis of carbohydrates is its nondestructive nature. The oligosaccharide/glycopeptide sample, after NMR analysis, can be recovered 100% unimpaired and used for other analyses, and biological activity tests. Also, mixtures of components with closely related structures can be analyzed successfully.