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Investigation Of The Saccharomyces Cerevisiae Gpi Transamidase: Insights Into Its Activity And Subunit-Subunit Interactions, Travis Ness
Wayne State University Dissertations
Glycosylphosphatidylinositol (GPI) anchoring of proteins is a eukaryotic, posttranslational
modification catalyzed by GPI transamidase (GPI-T). The Saccharomyces
cerevisiae GPI-T is composed of five membrane-bound subunits: Gaa1, Gpi8, Gpi16,
Gpi17, and Gab1. Structural and functional studies have been hindered by the
complexity of this enzyme. Conditions to purify the Gpi8:Gaa1:Gpi16 GPI-T heterotrimer
from yeast have been reported, but an understanding of the subunit functions,
interactions, and stoichiometry remain unclear. Furthermore, a reliable, quantitative, in
vitro assay for this important post-translational modification has remained elusive for
nearly three decades.
Our laboratory has developed an in vitro peptide cleavage assay that correlates
changes …