Holkeri H, Simonen M, Pummi T, Vihinen H, Makarow M
Institute of Biotechnology, University of Helsinki, Finland.
FEBS Lett. 1996 Apr 1;383(3):255-8. doi: 10.1016/0014-5793(96)00264-5.
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.
在此，我们研究了一种哺乳动物蛋白——大鼠神经生长因子受体（NGFRe）的胞外结构域在酿酒酵母中的糖基化情况。如果将NGFRe与一种多肽（hsp 150δ）载体融合，它会分泌到酿酒酵母的培养基中。hsp 150δ载体有95个丝氨酸和苏氨酸残基，这些残基被广泛地进行了O-糖基化。尽管有41个潜在位点，但无论是否与载体融合，NGFRe都缺乏O-聚糖。通过抑制二硫键形成来扭曲NGFRe的构象并不会促进O-糖基化，而N-糖基化则会增强。因此，hsp 150δ-NGFRe融合蛋白的丝氨酸和苏氨酸残基被高度选择性地进行了O-糖基化。
