Title : N-Glycosylation of secretion enhancer
peptide as influencing
factor for the secretion of target
proteins from Saccharomyces cerevisiae
Abstract :
- hIL-1beta-derived polypeptide , when fused to the N-terminal end of target proteins , exerts a potent secretion enhancer function in Saccharomyces cerevisiae
- We investigated the effect of N-glycosylation of the secretion enhancer peptide on the secretion of target proteins
- The N-terminal 24 amino acids (Ser5-Ala28) of human interleukin 1beta ( hIL-1beta ) and interleukin 1 receptor antagonist ( IL-1ra ) were used as secretion enhancer for synthesizing recombinant human granulocyte-colony stimulating factor ( rhG- CSF ) from S. cerevisiae
- The mutation of potential N-glycosylation site , by substituting Gln for either Asn7 of N-terminal 24 amino acids of hIL-1beta ( Asn7Gln ) or Asn84 of IL-1ra ( Asn84Gln ), resulted in a dramatic reduction of rhG- CSF secretion efficiency
- In contrast, the mutant containing an additional N-glycosylation site on the N-terminal 24 amino acids of hIL-1beta ( Gln15Asn ) secreted twice as much rhG- CSF into culture media as wild type hIL-1beta
- These results show that N-glycosylation of the secretion enhancer peptide plays an important role in increasing the secretion efficiency of the downstream target proteins
- The results also suggest that judicious choice of enhancer peptide and the control of its glycosylation could be of general utility for secretory production of heterologous proteins from S. cerevisiae