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Alpha‐Factor Pro‐Peptide N‐linked Oligosaccharides Facilitate Secretion of the Insulin Precursor in Saccharomyces Cerevisiae
Author(s) -
Kjeldsen Thomas,
Andersen Asser S.,
Hach Morten,
Diers Ivan,
Nikolajsen Jesper,
Markussen Jan
Publication year - 1998
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1998.tb01382.x
Subject(s) - glycosylation , secretion , biochemistry , saccharomyces cerevisiae , protein precursor , peptide , insulin , secretory pathway , biology , c peptide , n linked glycosylation , chemistry , glycoprotein , endocrinology , glycan , golgi apparatus , endoplasmic reticulum , yeast , gene
Toevaluate the possible relationship between N‐linked glycosylation ofthe Saccharomycescerevisiae a‐factor pro‐peptideand transport of the a‐factor pro‐peptide/insulin precursorfusion protein through the Saccharomyces cerevisiae secretory pathway, we analysed secretion of insulin precursor facilitated by a‐factor pro‐peptides with one or more of the three N‐linked glycosylation sites removed. Mutation of the three a‐factor pro‐peptide N‐linked glycosylation sites drastically decreased insulin precursor secretion. The three a‐factor pro‐peptide N‐linked glycosylation sites differ in their ability to facilitate secretion of the insulin precursor. The two a‐factor pro‐peptide N‐linked glycosylation sites localized closest to the insulin precursor contributed significantly to secretion, whereas the most N‐terminally linked glycosylation site did not appear to facilitate secretion. Only correctly folded insulin precursor was found in the culture supernatant, regardless of the pro‐peptide used for secretion, indicating that a‐factor pro‐peptide N‐linked oligosaccharide chains are not necessary for correct folding of the insulin precursor. Thus, N‐linked glycosylation facilitates intracellular transport of the a‐factor propeptide/insulin precursor fusion protein through the Saccharomyces cerevisiae secretory pathway and secretion of the insulin precursor. N‐linked glycosylation per se is not sufficient to facilitate secretion of the insulin precursor; the position of the N‐linked oligosaccharide chain on the a‐factor pro‐peptide is important for facilitating efficient secretion.