Disease mutations reveal residues critical to the interaction of P4-ATPases with lipid substrates

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  • Rasmus H Gantzel
  • Louise S Mogensen
  • ,
  • Stine A Mikkelsen
  • Bente Vilsen
  • Robert S Molday, Department of Ophthalmology and Visual Sciences, Centre for Macular Research, University of British Columbia, Vancouver, British Columbia, V6T 1Z3, Canada., Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, V6T 1Z3, Canada, Canada
  • Anna L Vestergaard, Laboratory for Immuno-Endocrinology, Department of Biomedical Sciences, University of Copenhagen, DK-2200, Copenhagen N, Denmark.
  • ,
  • Jens P Andersen

Phospholipid flippases (P4-ATPases) translocate specific phospholipids from the exoplasmic to the cytoplasmic leaflet of membranes. While there is good evidence that the overall molecular structure of flippases is similar to that of P-type ATPase ion-pumps, the transport pathway for the "giant" lipid substrate has not been determined. ATP8A2 is a flippase with selectivity toward phosphatidylserine (PS), possessing a net negatively charged head group, whereas ATP8B1 exhibits selectivity toward the electrically neutral phosphatidylcholine (PC). Setting out to elucidate the functional consequences of flippase disease mutations, we have identified residues of ATP8A2 that are critical to the interaction with the lipid substrate during the translocation process. Among the residues pinpointed are I91 and L308, which are positioned near proposed translocation routes through the protein. In addition we pinpoint two juxtaposed oppositely charged residues, E897 and R898, in the exoplasmic loop between transmembrane helices 5 and 6. The glutamate is conserved between PS and PC flippases, whereas the arginine is replaced by a negatively charged aspartate in ATP8B1. Our mutational analysis suggests that the glutamate repels the PS head group, whereas the arginine minimizes this repulsion in ATP8A2, thereby contributing to control the entry of the phospholipid substrate into the translocation pathway.

Original languageEnglish
Article numberdoi: 10.1038/s41598-017-10741-z.
JournalScientific Reports
Volume7
Issue1
Pages (from-to)10418
Number of pages11
ISSN2045-2322
DOIs
Publication statusPublished - 5 Sep 2017

    Research areas

  • Journal Article

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