Glycosylation of rat NGF receptor ectodomain in the yeast Saccharomyces cerevisiae.
Here we studied the glycosylation of a mammalian protein, the ectodomain of rat nerve growth factor receptor (NGFRe), in Saccharomyces cerevisiae. NGFRe is secreted to the culture medium of S. cerevisiae if it is fused to a polypeptide (hsp 150 delta) carrier. The hsp 150 delta-carrier has 95 serine and threonine residues, which were extensively O-glycosylated. In spite of 41 potential sites, NGFRe lacked O-glycans, whether fused to the carrier or not. Distortion of the conformation of NGFRe by inhibition of disulfide formation did not promote O-glycosylation, whereas N-glycosylation was enhanced. Thus, the serine and threonine residues of the hsp 150 delta-NGFRe fusion protein were highly selectively O-glycosylated.
Holkeri
H
H
Institute of Biotechnology, University of Helsinki, Finland.
Simonen
M
M
Pummi
T
T
Vihinen
H
H
Makarow
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001224:Aspartic Acid; D001345:Autoradiography; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003545:Cysteine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D006023:Glycoproteins; D006031:Glycosylation; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000378:metabolism; D008358:Mannose / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011487:Protein Conformation; D051381:Rats; D017475:Receptors, Nerve Growth Factor / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D013462:Sulfur Radioisotopes; D014316:Tritium
D006023:Glycoproteins; C074229:HSP150 protein, S cerevisiae; D006360:Heat-Shock Proteins; D009838:Oligodeoxyribonucleotides; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D013462:Sulfur Radioisotopes; D014316:Tritium; D001224:Aspartic Acid; D008715:Methionine; D003545:Cysteine; D008358:Mannose