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Anal Chem. 2015 Apr 21;87(8):4402-8. doi: 10.1021/acs.analchem.5b00170. Epub 2015 Apr 9.

Picodiscs for facile protein-glycolipid interaction analysis.

Author information

1
§Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
2
∥Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada M5G 1L7.

Abstract

Protein interactions with glycolipids are implicated in diverse cellular processes. However, the study of protein-glycolipid complexes remains a significant experimental challenge. Here, we describe a powerful new assay that combines electrospray ionization mass spectrometry (ESI-MS) and picodiscs, which are composed of human sphingolipid activator protein saposin A and a small number of phospholipids, to display glycolipids in a lipid environment for protein-glycolipid interaction studies in aqueous solution. Time-resolved measurements of enzyme catalyzed hydrolysis of glycolipid substrates and the detection of low, moderate, and high affinity protein-glycolipid interactions serve to demonstrate the reliability and versatility of the assay.

PMID:
25803566
DOI:
10.1021/acs.analchem.5b00170
[Indexed for MEDLINE]

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