Cryo-EM analysis of the conformational landscape of human P-glycoprotein (ABCB1) during its catalytic cycle

Gabriel A. Frank, Suneet Shukla, Prashant Rao, Mario J. Borgnia, Alberto Bartesaghi, Alan Merk, Aerfa Mobin, Lothar Esser, Lesley A. Earl, Michael M. Gottesman, Di Xia, Suresh V. Ambudkar, Sriram Subramaniam

Research output: Contribution to journalArticle

Abstract

The multidrug transporter P-glycoprotein (P-gp, ABCB1) is an ATP-dependent pump that mediates the efflux of structurally diverse drugs and xenobiotics across cell membranes, affecting drug pharmacokinetics and contributing to the development of multidrug resistance. Structural information about the conformational changes in human P-gp during the ATP hydrolysis cycle has not been directly demonstrated, although mechanistic information has been inferred from biochemical and biophysical studies conducted with P-gp and its orthologs, or from structures of other ATP-binding cassette transporters. Using single-particle cryo-electron microscopy, we report the surprising discovery that, in the absence of the transport substrate and nucleotides, human P-gp can exist in both open [nucleotide binding domains (NBDs) apart; inwardfacing] and closed (NBDs close; outward-facing) conformations. We also probe conformational states of human P-gp during the catalytic cycle, and demonstrate that, following ATP hydrolysis, P-gp transitions through a complete closed conformation to a complete open conformation in the presence of ADP.

Original languageEnglish (US)
Pages (from-to)35-41
Number of pages7
JournalMolecular Pharmacology
Volume90
Issue number1
DOIs
StatePublished - Jul 1 2016
Externally publishedYes

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P-Glycoprotein
Nucleotides
Adenosine Triphosphate
Hydrolysis
Cryoelectron Microscopy
ATP-Binding Cassette Transporters
Multiple Drug Resistance
Xenobiotics
Pharmaceutical Preparations
Adenosine Diphosphate
Pharmacokinetics
Cell Membrane

ASJC Scopus subject areas

  • Pharmacology
  • Molecular Medicine

Cite this

Frank, G. A., Shukla, S., Rao, P., Borgnia, M. J., Bartesaghi, A., Merk, A., ... Subramaniam, S. (2016). Cryo-EM analysis of the conformational landscape of human P-glycoprotein (ABCB1) during its catalytic cycle. Molecular Pharmacology, 90(1), 35-41. https://doi.org/10.1124/mol.116.104190

Cryo-EM analysis of the conformational landscape of human P-glycoprotein (ABCB1) during its catalytic cycle. / Frank, Gabriel A.; Shukla, Suneet; Rao, Prashant; Borgnia, Mario J.; Bartesaghi, Alberto; Merk, Alan; Mobin, Aerfa; Esser, Lothar; Earl, Lesley A.; Gottesman, Michael M.; Xia, Di; Ambudkar, Suresh V.; Subramaniam, Sriram.

In: Molecular Pharmacology, Vol. 90, No. 1, 01.07.2016, p. 35-41.

Research output: Contribution to journalArticle

Frank, GA, Shukla, S, Rao, P, Borgnia, MJ, Bartesaghi, A, Merk, A, Mobin, A, Esser, L, Earl, LA, Gottesman, MM, Xia, D, Ambudkar, SV & Subramaniam, S 2016, 'Cryo-EM analysis of the conformational landscape of human P-glycoprotein (ABCB1) during its catalytic cycle', Molecular Pharmacology, vol. 90, no. 1, pp. 35-41. https://doi.org/10.1124/mol.116.104190
Frank, Gabriel A. ; Shukla, Suneet ; Rao, Prashant ; Borgnia, Mario J. ; Bartesaghi, Alberto ; Merk, Alan ; Mobin, Aerfa ; Esser, Lothar ; Earl, Lesley A. ; Gottesman, Michael M. ; Xia, Di ; Ambudkar, Suresh V. ; Subramaniam, Sriram. / Cryo-EM analysis of the conformational landscape of human P-glycoprotein (ABCB1) during its catalytic cycle. In: Molecular Pharmacology. 2016 ; Vol. 90, No. 1. pp. 35-41.
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