title: Picodiscs for facile protein-glycolipid interaction analysis.
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.
authors: Aneika C
authors: AC
authors: Leney
authors: Reza
authors: R
authors: Rezaei Darestani
authors: Jun
authors: J
authors: Li
authors: Sanaz
authors: S
authors: Nikjah
authors: Elena N
authors: EN
authors: Kitova
authors: Chunxia
authors: C
authors: Zou
authors: Christopher W
authors: CW
authors: Cairo
authors: §Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
authors: Zi Jian
authors: ZJ
authors: Xiong
authors: §Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8.|∥Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada M5G 1L7.
authors: Gilbert G
authors: GG
authors: Privé
authors: John S
authors: JS
authors: Klassen
mesh_terms: D006017:Glycolipids / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D009439:Neuraminidase / Q000737:chemistry / Q000378:metabolism; D049231:Saposins / Q000737:chemistry* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors
chemical_list: D006017:Glycolipids; D049231:Saposins; C465436:Neu3 protein, human; D009439:Neuraminidase