title: The position of the oligosaccharide side-chains of phytohemagglutinin and their accessibility to glycosidases determines their subsequent processing in the Golgi.
abstract: Phytohemagglutinin (PHA), the glycoprotein lectin of Phaseolus vulgaris has two types of asparagine-linked oligosaccharides per polypeptide: a high-mannose chain with the formula (Man)8-9(GlcNAc)2 on Asn12 and a modified chain with fewer mannose residues and additional fucose and xylose residues on Asn60. Glycosylation of PHA is a cotranslational process, which occurs in the endoplasmic reticulum, and newly synthesized PHA has two high-mannose chains. Transport of PHA to the protein bodies via the Golgi complex is accompanied by the modification of one of the two high-mannose chains. Why is only one chain modified, while the other remains in the high-mannose configuration? By determining the effect of digestion with various glycosidases (alpha-mannosidase, endo-beta-N-acetylglucosaminidase H and endo-beta-N-acetylglucosaminidase F) on native and denatured PHA we obtained evidence consistent with the interpretation that the accessibility of oligosaccharide chains to modifying enzymes is of major importance in determining whether a high-mannose chain becomes modified or not. The high-mannose chain of mature undenatured PHA is only partially accessible to glycosidases, while PHA obtained from the endoplasmic reticulum has one high-mannose chain, which is readily accessible to alpha-mannosidase and endoglycosidases H and F. We show that this readily accessible chain is in the same position on the polypeptide (Asn60) as the modified oligosaccharide on mature PHA. Thus, accessibility of the oligosaccharide side-chains to processing enzymes in the Golgi determines whether a particular oligosaccharide side-chain is processed or not.
mesh_terms: D003208:Concanavalin A / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000032:analysis; D005643:Fucose / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000494:pharmacology*; D006056:Golgi Apparatus / Q000378:metabolism*; D006596:Hexosaminidases / Q000494:pharmacology; D008361:Mannosidases / Q000494:pharmacology; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D010835:Phytohemagglutinins / Q000378:metabolism*; D011489:Protein Denaturation; D043323:alpha-Mannosidase
chemical_list: D009844:Oligosaccharides; D010835:Phytohemagglutinins; D003208:Concanavalin A; D005643:Fucose; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; D008361:Mannosidases; D043323:alpha-Mannosidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D005944:Glucosamine