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CFTR Folding Consortium: Methods Available for Studies of CFTR Folding and Correction

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Cystic Fibrosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 742))

Abstract

The CFTR Folding Consortium (CFC) was formed in 2004 under the auspices of the Cystic Fibrosis Foundation and its drug discovery and development affiliate, CFF Therapeutics. A primary goal of the CFC is the development and distribution of reagents and assay methods designed to better understand the mechanistic basis of mutant CFTR misfolding and to identify targets whose manipulation may correct CFTR folding defects. As such, reagents available from the CFC primarily target wild-type CFTR NBD1 and its common variant, F508del, and they include antibodies, cell lines, constructs, and proteins. These reagents are summarized here, and two protocols are described for the detection of cell surface CFTR: (a) an assay of the density of expressed HA-tagged CFTR by ELISA and (b) the generation and use of an antibody to CFTR’s first extracellular loop for the detection of endogenous CFTR. Finally, we highlight a systematic collection of assays, the CFC Roadmap, which is being used to assess the cellular locus and mechanism of mutant CFTR correction. The Roadmap queries CFTR structure–function relations at levels ranging from purified protein to well-differentiated human airway primary cultures.

K.W. Peters and T. Okiyoneda contributed equally to this chapter

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Acknowledgments

Resources providing support for this work in the Frizzell lab include grants from the NIH (DK068196 and DK 072506) and the Cystic Fibrosis Foundation (CFF R883-CR07 and FRIZZE05XX0). Experimental work in the laboratory of Gergely Lukacs was funded by the NIH, Cystic Fibrosis Folding Consortium, CIHR, and CFI. Tsukasa Okiyoneda was supported by a postdoctoral fellowship from the Canadian Cystic Fibrosis Foundation.

Author information

Authors and Affiliations

  1. Department of Cell Biology and Physiology, University of Pittsburgh, 15261, Pittsburgh, PA, USA

    Kathryn W. Peters

  2. Department of Physiology, McGill University, H3G 1Y6, Montreal, QC, Canada

    Tsukasa Okiyoneda & Gergely L. Lukacs

  3. Department of Cell Biology, The Scripps Research Institute, 92037, La Jolla, CA, USA

    William E. Balch

  4. Cellular Protein Chemistry, Utrecht University, Utrecht, The Netherlands

    Ineke Braakman

  5. Department of Biological Sciences, University of Pittsburgh, 15260, Pittsburgh, PA, USA

    Jeffrey L. Brodsky

  6. Department of Physiology, Johns Hopkins University, 21205, Baltimore, MD, USA

    William B. Guggino

  7. Cystic Fibrosis Foundation, 20814, Bethesda, MD, USA

    Christopher M. Penland

  8. Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, 20814, Bethesda, MD, USA

    Harvey B. Pollard

  9. Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, 35294, Birmingham, AL, USA

    Eric J. Sorscher

  10. Department of Biochemistry and Molecular Biology, Oregon Health & Science University, 97229, Portland, OR, USA

    William R. Skach

  11. Department of Physiology, University of Texas Southwestern Medical Center, 75390, Dallas, TX, USA

    Philip J. Thomas

  12. Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, 15261, Pittsburgh, PA, USA

    Raymond A. Frizzell

Authors
  1. Kathryn W. Peters
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  2. Tsukasa Okiyoneda
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  3. William E. Balch
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  4. Ineke Braakman
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  5. Jeffrey L. Brodsky
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  6. William B. Guggino
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  7. Christopher M. Penland
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  8. Harvey B. Pollard
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  9. Eric J. Sorscher
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  10. William R. Skach
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  11. Philip J. Thomas
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  12. Gergely L. Lukacs
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  13. Raymond A. Frizzell
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Corresponding author

Correspondence to Raymond A. Frizzell .

Editor information

Editors and Affiliations

  1. Faculty of Sciences, Centre for Biodiversity & Functional, University of Lisboa, Campo Grande, Lisboa, 1749-016, Portugal

    Margarida D. Amaral

  2. , Department of Physiology, University of Regensburg, Universitätsstr. 31, Regensburg, 93053, Germany

    Karl Kunzelmann

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Peters, K.W. et al. (2011). CFTR Folding Consortium: Methods Available for Studies of CFTR Folding and Correction. In: Amaral, M., Kunzelmann, K. (eds) Cystic Fibrosis. Methods in Molecular Biology, vol 742. Humana Press. https://doi.org/10.1007/978-1-61779-120-8_20

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  • DOI: https://doi.org/10.1007/978-1-61779-120-8_20

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-119-2

  • Online ISBN: 978-1-61779-120-8

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