Biosynthesis and processing of phytohemagglutinin in developing bean cotyledons.
Phytohemagglutinin (PHA) is a family of tetrameric isolectins which accumulate in the protein bodies of developing Phaseolus vulgaris cotyledons. Each tetramer contains erythroagglutinating (E) or lymphocyte-mitogenic (L) subunits, or a combination of both. The subunits have Mr around 33000, E being slightly larger than L. Phytohemagglutinin is a glycoprotein, and its carbohydrate moiety contains N-acetylglucosamine, mannose, fucose and xylose, indicating that this protein has complex oligosaccharide sidechains. Several steps in the biosynthesis and in the cotranslational and post-translational processing of the glycopolypeptides of PHA have been identified. The polypeptides of PHA are synthesized by polysomes attached to the endoplasmic reticulum. The glycosylation of the polypeptides is a cotranslational process, in which each PHA polypeptide usually acquires two oligosaccharide sidechains. The oligosaccharides of PHA isolated from the endoplasmic reticulum are susceptible to digestion with alpha-mannosidase and endo-beta-N-acetylglucosaminidase H indicating that they are of the high-mannose type. In the presence of tunicamycin two unglycosylated polypeptides of PHA are synthesized, indicating that the differences in Mr between the E and L subunits of PHA are not due to differences in glycosylation alone. Transport of PHA to the protein bodies is mediated by the Golgi apparatus where at least part of the oligosaccharide chains of PHA are modified [ Chrispeels , M. J. (1983) Planta ( Berl .) 157, 454-461, and 158, 140-151]. The modified oligosaccharide chains of PHA are then gradually trimmed to a smaller size when the protein is already in the protein bodies. This processing results in an increase in the mobility of the PHA subunits in denaturing polyacrylamide gels.
D004721:Endoplasmic Reticulum / Q000378:metabolism; D007887:Fabaceae / Q000254:growth & development / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D010835:Phytohemagglutinins / Q000096:biosynthesis*; D037121:Plant Lectins; D010946:Plants, Medicinal; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D013050:Spectrometry, Fluorescence; D014415:Tunicamycin / Q000494:pharmacology
D009844:Oligosaccharides; D010835:Phytohemagglutinins; D037121:Plant Lectins; D014415:Tunicamycin; D008358:Mannose