De Novo Missense Variations of ATP8B2 Impair Its Phosphatidylcholine Flippase Activity

Mol Cell Biol. 2024;44(11):473-488. doi: 10.1080/10985549.2024.2391829. Epub 2024 Sep 2.

Abstract

P4-ATPases comprise a family of lipid flippases that translocate lipids from the exoplasmic (or luminal) to the cytoplasmic leaflet of biological membranes. Of the 14 known human P4-ATPases, ATP8B2 is a phosphatidylcholine flippase at the plasma membrane, but its physiological function is not well understood. Although ATP8B2 could interact with both CDC50A and CDC50B, it required only the CDC50A interaction for its exit from the endoplasmic reticulum and subsequent transport to the plasma membrane. Three de novo monoallelic missense variations of ATP8B2 were found in patients with intellectual disability. None of these variations affected the interaction of ATP8B2 with CDC50A or its localization to the plasma membrane. However, variations of either of two amino acid residues, which are conserved in all P4-ATPases, significantly reduced the phosphatidylcholine flippase activity of ATP8B2. Furthermore, mutations in the corresponding residues of ATP8B1 and ATP11C were found to decrease their flippase activities toward phosphatidylcholine and phosphatidylserine, respectively. These results indicate that the conserved amino acid residues are crucial for the enzymatic activities of the P4-ATPases.

Keywords: Lipid bilayer; P-type ATPase; flippase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases* / genetics
  • Adenosine Triphosphatases* / metabolism
  • Amino Acid Sequence
  • Cell Membrane / metabolism
  • Endoplasmic Reticulum / metabolism
  • HEK293 Cells
  • Humans
  • Intellectual Disability / genetics
  • Intellectual Disability / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Membrane Transport Proteins
  • Mutation, Missense*
  • Phosphatidylcholines* / metabolism
  • Phosphatidylserines / metabolism
  • Phospholipid Transfer Proteins / genetics
  • Phospholipid Transfer Proteins / metabolism

Substances

  • Adenosine Triphosphatases
  • Phosphatidylcholines
  • ATP8A2 protein, human
  • Phospholipid Transfer Proteins
  • ATP11C protein, human
  • TMEM30a protein, human
  • Membrane Proteins
  • Phosphatidylserines
  • ATP8B1 protein, human
  • Membrane Transport Proteins

Grants and funding

This work was supported by the JSPS KAKENHI under Grant JP23H02434, JP23K27127, and JP20H03209 (to H.-W.S.) and JP20K07325 (to H.T.); the Takeda Science Foundation (to H.-W.S.); the Uehara Memorial Foundation (to H.-W.S.); the Mizutani Glycoscience Foundation (to H.-W.S.); and the ONO Medical Research Foundation (to H.-W.S.).