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Membrane transplantation to correct integral membrane protein defects

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Abstract

In this report we show that the tendency of certain viruses to carry host membrane proteins in their envelopes can be harnessed for transplantation of small patches of plasma membrane, including fully functional, polytopic ion channel proteins and their regulatory binding partners. As a stringent model we tested the topologically complex epithelial ion channel CFTR. Initially an attenuated vaccinia virus was found capable of transferring CFTR in a properly folded, functional and regulatable form to CFTR negative cells. Next we generated viruslike particles (VLPs) composed of retroviral structural proteins that assemble and bud at the host cell plasma membrane. These particles were also shown to mediate functional ion channel transfer. By testing the capacity of complex membrane proteins to incorporate into viral envelopes these experiments provide new insight into the permissiveness of viral envelopment, including the ability of incorporated proteins to retain function and repair defects at the cell surface, and serve as a platform for studies of ion channel and membrane protein biochemistry.

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Abbreviations

CFTR :

Cystic fibrosis transmembrane conductance regulator

EMV :

Extracellular mature virus

ER :

Endoplasmic reticulum

IMV :

Intracellular mature virus

MOI :

Multiplicity of infection

PKA :

Protein kinase A

RSV :

Rous sarcoma virus

VLP :

Viruslike particle

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Acknowledgements

We thank Drs. Chris Yun and Mark Donowitz (Johns Hopkins University) for providing E3KARP antibody, Drs. Kevin Kirk and Estelle Cormet-Boyaka (University of Alabama at Birmingham) for providing SNAP-23 antibody, and Dr. Min Li (Johns Hopkins University) for antibody to CAP70. We also thank John Wills for pMyr1a construct, Eddie Walthall, Zsuzsa Bebok, and Susan Dubay for assistance and discussions, Leigh Millican at the University of Alabama at Birmingham Electron Microscopy Core, and Mrs. Mikelle Foster for help in manuscript preparation. This work is supported by the NIH and CFF.

Author information

Author notes

  1. Scott A. King

    Present address: Department of Physiology, School of Medicine, University of California at Davis, One Shields Avenue, CA 95616-8644, Davis, USA

  2. Michael Sakalian

    Present address: Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73190, USA

Authors and Affiliations

  1. Department of Human Genetics, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Kimberly V. Curlee

  2. Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Jeong S. Hong

  3. Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    J. P. Clancy

  4. Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, 1530 3rd Avenue South, MCLM 796, Birmingham, AL, 35294-0005, USA

    Kimberly V. Curlee, Jeong S. Hong, J. P. Clancy, Scott A. King & Eric J. Sorscher

  5. Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Scott A. King, Bakhrom Berdiev, Dale Benos & Eric J. Sorscher

  6. Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Eric J. Sorscher

  7. Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Eric Hunter & Michael Sakalian

  8. Centre for Research in Virology, Department of Microbiology and Immunology, Gade Institute, University of Bergen, Bergen High Technology Centre, 5020, Bergen , Norway

    Maja A. Sommerfelt

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  1. Kimberly V. Curlee
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Corresponding author

Correspondence to Eric J. Sorscher.

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Curlee, K.V., Hong, J.S., Clancy, J.P. et al. Membrane transplantation to correct integral membrane protein defects. J Mol Med 81, 511–520 (2003). https://doi.org/10.1007/s00109-003-0446-7

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  • DOI: https://doi.org/10.1007/s00109-003-0446-7

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