Biosynthesis and Characterization of Early Glycosylphosphatidylinositol Anchor Intermediates in Saccharomyces Cerevisiae

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.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Costello, Lisa Catherine

Contributors dc:contributor

• Peter A.B.Orlean

• Biology, Molecular

Language dc:language

eng

Identifier

(MiAaPQ)AAI9912214

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/84896