Title: | Glycosyl-phosphatidylinositol anchor attachment in a yeast in vitro system |
Address: | "Department of Molecular and Cell Biology, Barker Hall, Howard Hughes Research Institute, University of California, Berkeley, CA 94720-3202, USA" |
ISSN/ISBN: | 0264-6021 (Print) 1470-8728 (Electronic) 0264-6021 (Linking) |
Abstract: | "The yeast mating pheromone precursor prepro-alpha factor was fused to C-terminal signals for glycosyl-phosphatidylinositol (GPI) anchor attachment, based on the sequence of the Saccharomyces cerevisiae protein Gas1p. Maturation of fusion proteins expressed in vivo required the presence of both a functional GPI attachment site and the synthesis of GPI precursors. Constructs were translated in vitro for use in cell-free studies of glycolipid attachment. The radiolabelled polypeptides were post-translationally translocated into yeast microsomes, where at least one third of the molecules received a GPI anchor. This approach offers distinct advantages over anchor attachment reactions that require co-translational translocation of secretory peptide substrates" |
Keywords: | Biological Transport Cell-Free System Fungal Proteins/genetics/*metabolism Glycosylphosphatidylinositols/*metabolism Membrane Glycoproteins/genetics/*metabolism Protein Precursors/genetics/*metabolism Recombinant Fusion Proteins/metabolism Research Design; |
Notes: | "MedlineDoering, T L Schekman, R eng Research Support, Non-U.S. Gov't England 1998/01/24 Biochem J. 1997 Dec 1; 328 ( Pt 2)(Pt 2):669-75. doi: 10.1042/bj3280669" |