CFTR, the odd ABC transporter responsible for cystic fibrosis Jue - - PowerPoint PPT Presentation

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CFTR, the odd ABC transporter responsible for cystic fibrosis Jue - - PowerPoint PPT Presentation

Dec 29, 2022 288 likes •677 views

CFTR, the odd ABC transporter responsible for cystic fibrosis Jue Chen Laboratory of Membrane Biology and Biophysics Cystic Fibrosis is a genetic disease cftr Picture credit: Bruce Blaus CFTR, an odd ABC transporter Typical ABC transporters:

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Jue Chen Laboratory of Membrane Biology and Biophysics

CFTR, the odd ABC transporter responsible for cystic fibrosis

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Picture credit: Bruce Blaus

cftr

Cystic Fibrosis is a genetic disease

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CFTR, an odd ABC transporter

Typical ABC transporters: ATP-powered pumps CFTR: an ATP-gated anion channel

Phosphorylation

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Phosphorylation gates the CFTR channel

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Phosphorylation also regulates ATPase activity

Zhe Zhang

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Cryo-EM analysis of CFTR

Zhe Zhang

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Cryo-EM analysis of CFTR

  • Movie alignment and exposure filter
  • - Unblur: Grant & Grigorieff, eLIFE, 2015
  • Particle alignment
  • - alignparts_lmbfgs: Rubinstein & Brubaker, J Struct Biol. 2014
  • Estimate defocus parameters
  • - CTFFND4: Rohou & Grigorieff, J Struct Biol. (2015)
  • Particle picking and subsequent processing
  • - Relion: Scheres, J Struct Biol. 2015
  • Final runs of 3D refinement
  • - Frealign: Lyumkis & Grigorieff, J Struct Biol. 2013
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Cryo-EM analysis of zebrafish CFTR

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Cryo-EM analysis of zebrafish CFTR

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Cryo-EM analysis of zebrafish CFTR

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Zebrafish CFTR

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Human CFTR

Fangyu Liu and Zhe Zhang

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The ion conduction pathway

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Positively charged residues

Reviewed in Linsdell, 2017

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Anion selectivity

/e /e

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Selectivity

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David C. Gadsby

Towards an open channel conformation

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Cryo-EM analysis of phosphorylated CFTR

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Cryo-EM analysis of phosphorylated CFTR

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Cryo-EM analysis of phosphorylated CFTR

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Cryo-EM analysis of phosphorylated CFTR

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R domain prevents NBD dimerization

Dephosphorylated ATP-free

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R domain prevents NBD dimerization

Phosphorylated ATP-bound

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Conformational changes

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Conformational changes

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Local conformations control ion access

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David C. Gadsby

Flickering

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The CFTR gating cycle

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The CFTR gating cycle

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Cystic Fibrosis-causing mutations

  • >1000 mutations in CFTR
  • 272 mutations causing CF
  • 53 missense mutations at 46 positions
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Cystic Fibrosis-causing mutations

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Pore construction mutants

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Folding mutants

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ATPase site mutants

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NBD/TMD interface mutants

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NBD/TMD interface mutants

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Cystic Fibrosis-causing mutations

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Acknowledgment

EM Mentors

  • Tom Walz
  • Zongli Li
  • Rich Hite
  • Nikolaus Grigorieff
  • Tim Grant
  • Alexis Rohou

Staff at RU and Janelia

  • Mark Ebrahim
  • Johanna Sotiris
  • Zhiheng Yu
  • Chuang Hong
  • Rick Huang

CFTR project Zhe Zhang Fangyu Liu David C. Gadsby