Biosynthesis and Characterization of Early Glycosylphosphatidylinositol Anchor Intermediates in Saccharomyces Cerevisiae
Costello, Lisa Catherine
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https://hdl.handle.net/2142/84896
Description
Title
Biosynthesis and Characterization of Early Glycosylphosphatidylinositol Anchor Intermediates in Saccharomyces Cerevisiae
Author(s)
Costello, Lisa Catherine
Issue Date
1998
Doctoral Committee Chair(s)
Peter A.B.Orlean
Department of Study
Biochemistry
Discipline
Biochemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Language
eng
Abstract
Next, it was desired to obtain structural information on in vivo GPI anchor intermediates. To address this issue, I exploited the fact that at non-permissive temperature, the yeast dpm1--6 mutant accumulates GlcNH2-(acyl-inositol)-Pl. This precursor molecule is the earliest biosynthetic intermediate that can be detected in in vivo radiolabeling experiments. I isolated this lipid and had it analyzed by liquid chromatography - mass spectroscopy (LC-MS) with an electrospray interface. The results show that this intermediate has very long chain fatty acids, most likely on its diacylglycerol moiety. This has significant implications for GPI anchor biosynthesis in yeast and suggests that GPI anchors are either assembled on a specialized PI or that the diacylglycerol moiety on early GPI precursor molecules are rapidly remodeled.
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