Structure of the human lipid exporter ABCB4 in a lipid environment

Nat Struct Mol Biol. 2020 Jan;27(1):62-70. doi: 10.1038/s41594-019-0354-3. Epub 2019 Dec 23.

Abstract

ABCB4 is an ATP-binding cassette transporter that extrudes phosphatidylcholine into the bile canaliculi of the liver. Its dysfunction or inhibition by drugs can cause severe, chronic liver disease or drug-induced liver injury. We determined the cryo-EM structure of nanodisc-reconstituted human ABCB4 trapped in an ATP-bound state at a resolution of 3.2 Å. The nucleotide binding domains form a closed conformation containing two bound ATP molecules, but only one of the ATPase sites contains bound Mg2+. The transmembrane domains adopt a collapsed conformation at the level of the lipid bilayer, but we observed a large, hydrophilic and fully occluded cavity at the level of the cytoplasmic membrane boundary, with no ligand bound. This indicates a state following substrate release but prior to ATP hydrolysis. Our results rationalize disease-causing mutations in human ABCB4 and suggest an 'alternating access' mechanism of lipid extrusion, distinct from the 'credit card swipe' model of other lipid transporters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B / chemistry*
  • ATP Binding Cassette Transporter, Subfamily B / metabolism
  • ATP Binding Cassette Transporter, Subfamily B / ultrastructure
  • Adenosine Triphosphate / metabolism
  • Binding Sites
  • Cryoelectron Microscopy
  • Humans
  • Hydrolysis
  • Lipid Bilayers / chemistry
  • Models, Molecular
  • Protein Conformation
  • Substrate Specificity

Substances

  • ATP Binding Cassette Transporter, Subfamily B
  • Lipid Bilayers
  • Adenosine Triphosphate
  • multidrug resistance protein 3