title: Enhanced secretion of biologically active, non-glycosylated VEGF from Saccharomyces cerevisiae.
abstract: Vascular endothelial growth factor (VEGF) mediates angiogenesis, which plays a critical role in the development and differentiation of the vascular system. VEGF is a homodimeric glycoprotein that contains one N-glycosylation site. In this study, we evaluated Saccharomyces cerevisiae expression systems producing glycosylated and non-glycosylated splice variants of human VEGF, VEGF121, and VEGF165. The pre region of the mating factor α1 (MFα1) signal sequence was found to perform better than the entire MFα1 prepro signal sequence in secreting glycosylated VEGF. Secretion of non-glycosylated VEGF165 was completely blocked, indicating the importance of glycosylation in VEGF165 secretion. Interestingly, non-glycosylated VEGF165 was secreted when guided by the MFα1 prepro signal sequence, albeit to a lesser degree, compared to glycosylated VEGF165. N-glycosylation in the pro region was required for the prepro sequence to promote VEGF secretion. Furthermore, substitution of asparagine at the VEGF glycosylation site with lysine or glutamic acid increased secretion of non-glycosylated VEGF, a finding not previously reported. Our findings suggest that S. cerevisiae could be a suitable host for secreting biologically active, non-glycosylated VEGF for clinical use.
authors: Department of Microbiology and Molecular Biology, College of Bioscience and Biotechnology, Chungnam National University, Daejeon 305-764, Republic of Korea.
authors: Woo Kyu
authors: WK
authors: Kang
authors: Min Hyung
authors: MH
authors: Lee
authors: Yeong Hyeock
authors: YH
authors: Kim
authors: Min Young
authors: MY
authors: Kim
authors: Jeong-Yoon
authors: JY
authors: Kim
mesh_terms: D001216:Asparagine; D001709:Biotechnology; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D010447:Peptide Hydrolases / Q000378:metabolism; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D001216:Asparagine; D010447:Peptide Hydrolases