Transmembrane polar relay drives the allosteric regulation for ABCG5/G8 sterol transporter

Bala M. Xavier, Aiman A. Zein, Angelica Venes, Junmei Wang, Jyh Yeuan Lee

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

The heterodimeric ATP-binding cassette (ABC) sterol transporter, ABCG5/G8, is responsible for the biliary and transintestinal secretion of cholesterol and dietary plant sterols. Missense mutations of ABCG5/G8 can cause sitosterolemia, a loss-of-function disorder characterized by plant sterol accumulation and premature atherosclerosis. A new molecular framework was recently established by a crystal structure of human ABCG5/G8 and reveals a network of polar and charged amino acids in the core of the transmembrane domains, namely, a polar relay. In this study, we utilize genetic variants to dissect the mechanistic role of this transmembrane polar relay in controlling ABCG5/G8 function. We demonstrated a sterol-coupled ATPase activity of ABCG5/G8 by cholesteryl hemisuccinate (CHS), a relatively water-soluble cholesterol memetic, and characterized CHS-coupled ATPase activity of three loss-of-function missense variants, R543S, E146Q, and A540F, which are respectively within, in contact with, and distant from the polar relay. The results established an in vitro phenotype of the loss-of-function and missense mutations of ABCG5/G8, showing significantly impaired ATPase activity and loss of energy sufficient to weaken the signal transmission from the transmembrane domains. Our data provide a biochemical evidence underlying the importance of the polar relay and its network in regulating the catalytic activity of ABCG5/G8 sterol transporter.

Original languageEnglish (US)
Article number8747
Pages (from-to)1-20
Number of pages20
JournalInternational journal of molecular sciences
Volume21
Issue number22
DOIs
StatePublished - Nov 2 2020
Externally publishedYes

Keywords

  • ABCG5
  • ABCG8
  • ATP-binding cassette transporter
  • Cholesterol
  • Polar relay
  • Sitosterolemia

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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