Cell surface glycoproteins undergo postbiosynthetic modification of their N-glycans by stepwise demannosylation.

The Journal of Biological Chemistry
S PorwollR Tauber

Abstract

Primary rat hepatocytes and two hepatoma cell lines have been used to study whether high mannose-type N-glycans of plasma membrane glycoproteins may be modified by the removal of mannose residues even after transport to the cell surface. To examine glycan remodeling of cell surface glycoproteins, high mannose-type glycoforms were generated by adding the reversible mannosidase I inhibitor deoxymannojirimycin during metabolic labeling with [3H]mannose, thereby preventing further processing of high mannose-type N-glycans to complex structures. Upon transport to the cell surface, glycoproteins were additionally labeled with sulfosuccinimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate. This strategy allowed us to follow selectively the fate of cell surface glycoproteins. Postbiosynthetic demannosylation was monitored by determining the conversion of Man8-9GlcNAc2 to smaller structures during reculture of cells in the absence of deoxymannojirimycin. The results show that high mannose-type N-glycans of selected cell surface glycoproteins are trimmed from Man8-9GlcNAc2 to Man5GlcNAc2 with Man7GlcNAc2 and Man6GlcNAc2 formed as intermediates. It could be clearly shown in MH 7777 as well as in HepG2 cells that demannosylation affects plasm...Continue Reading

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Citations

Jun 17, 2011·Biochemistry·Mahendra S SandbhorChristopher W Cairo
Dec 9, 2009·ACS Chemical Biology·Randy B Parker, Jennifer J Kohler
Jun 6, 2013·Biotechnology and Bioengineering·Rhian K Grainger, David C James
Jul 21, 2016·Angewandte Chemie·Paul R WratilWerner Reutter
Oct 12, 2018·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Guowei ZhangJianguo Gu
Jul 15, 2015·The Journal of Biological Chemistry·Jeffery ChenShetuan Zhang
Apr 16, 2010·Current Protocols in Protein Science·Giuliano Elia
May 22, 2001·Archives of Biochemistry and Biophysics·G OrbergerR Tauber

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