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Glycoprotein Analysis

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Molecular Biomethods Handbook

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

In eukaryottc cells, one of the most important posttranslattonal modrficattons of proteins is the co valent addmon of carbohydrate We can consider two major types of modtf-icatron to ammo-acid residues. N-glycosylation of asparagine amme groups and O-glycosylatron of serme or threonme hydroxyl groups (1). N-linked oltgosaccharrdes can be divided mto three maJor classes, the complex type contaming N-acetyl- glucosamme, mannose, galactose, fucose, and stahc acid; the oligomannose type contaming N-acetylglucosamme and mannose only; and the hybrid type that has features common to both complex and olrgomannose chains (Fig. 1) All of these structures are synthesized by a common pathway that begins in the endoplasmtc rettculum (ER) with the assembly of a lipid-lmked donor molecule The preformed oltgosacchartde is transferred to protein cotranslationally in the lumen of the endoplasmlc retrculum and by a serves of glycosrdase (a-glucosrdase and a-mannosrdase)-trmnning reactions is modr- fied as the protein progresses through the ER and Golgi apparatus (2), The diversity of N+lmked ohgosacchartde structure is dictated by the accesstbtllty of these partially processed chains to Golgi resident glycosyltransferases, a group of enzymes able to add monosacchartdes to oltgosacchartdes directly from nucleottde sugar donors. Glycosyltransferases are specrfic for nucleotide sugar donor, anomertctty, glycosrdtc linkage between sugars, and acceptor substrates. Consequently, there are a number of different transferases and each cell, tissue, and species has a unique complement of enzymes that control oltgosaccharrde biosynthesis (3). O-linked ohgosacchartdes con- tain similar residues to N-glycans, but their synthesis has no requirement for en bloc addttton of carbohydrate to the polypepttde chain O-glycosylatron proceeds by glycosyltransferase-catalyzed, stepwtse addition of monosacchartdes to generate, as in thecase of mucm glycoprotems, a diverse number of branched oltgosaccharrdes (4,5) (see Fig. 2).

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Authors and Affiliations

  1. Department of Btochemutry, Untverstty of Oxford, UK

    Terry D. Butters

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  1. Terry D. Butters
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  1. University of Herfordshire, Hatfield, UK

    Ralph Rapley  & John M. Walker  & 

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© 1998 Humana Press Inc , Totowa, NJ.

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Butters, T.D. (1998). Glycoprotein Analysis. In: Rapley, R., Walker, J.M. (eds) Molecular Biomethods Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-59259-642-3_40

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  • DOI: https://doi.org/10.1007/978-1-59259-642-3_40

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-501-0

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