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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Published on: April 9, 2018

Labeling cell surface glycosylphosphatidylinositol-anchored proteins through metabolic engineering using an

Sayan Kundu¹, Mohit Jaiswal¹, Kendall C Craig¹

¹Department of Chemistry, University of Florida, Gainesville, FL, 32611, USA.

Biochemical and Biophysical Research Communications

|January 22, 2023

Abstract:

Glycosylphosphatidylinositol (GPI) anchorage is one of the most common mechanisms to attach proteins to the plasma membrane of eukaryotic cells. GPI-anchored proteins (GPI-APs) play a critical role in many biological processes but are difficult to study. Here, a new method was developed for the effective and selective metabolic engineering and labeling of cell surface GPI-APs with an azide-modified phosphatidylinositol (PI) as the biosynthetic precursor of GPIs. It was demonstrated that this azido-PI derivative was taken up by HeLa cells and incorporated into the biosynthetic pathway of GPIs to present azide-labeled GPI-APs on the live cell surface. The azido group was used as a molecular handle to install other labels through a biocompatible click reaction to enable various biological studies, e.g., fluorescent imaging and protein pull-down, which can help explore the functions of GPI-APs and discover new GPI-APs.

Keywords:

Azide-modified phosphatidylinositol Glycosylphosphatidylinositol Glycosylphosphatidylinositol-anchored protein Metabolic engineering Phosphatidylinositol

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