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Permanent link (DOI): https://doi.org/10.7939/R39S1M01W
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Picodiscs for facile protein-glycolipid interaction analysis Open Access
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Leney, Aneika C.
Darestani, Reza Rezaei
Kitova, Elena N.
Cairo, Christopher W.Xiong, Zi Jian
Privé, Gilbert G.
Klassen, John S.
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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.
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- © 2015 Leney, A. C., Darestani, R. R., Li, J., Nikjah, S., Kitova, E. N., Zou, C., Cairo, C. W., Xiong, Z. J., Prive, G. G., & Klassen, J. S. This version of this article is open access and can be downloaded and shared. The original author(s) and source must be cited.
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Leney, A. C., Darestani, R. R., Li, J., Nikjah, S., Kitova, E. N., Zou, C., Cairo, C. W., Xiong, Z. J., Prive, G. G., & Klassen, J. S. (2015). Picodiscs for facile protein-glycolipid interaction analysis. Analytical Chemistry, 87(8), 4402-4408. http://doi.org/10.1021/acs.analchem.5b00170
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