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Biotin Identification Proteomics in Three-Dimensional Organotypic Human Skin Cultures

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Epidermal Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2109))

Abstract

Biotin identification (BioID) proteomics facilitates the unbiased detection of protein interaction neighborhoods in live cells. The BioID technique relies on the covalent biotin alteration of vicinal proteins by a modified bacterial biotin ligase. The biotin ligase is fused to a protein of interest to identify putative protein-protein interactions. Here, we describe the adaptation of this technique for use in three-dimensional epidermal cultures. Due to the covalent biotin modification of proteins, our protocol allows for the complete solubilization of the total cellular protein content in differentiated keratinocytes. Thus, a comprehensive network of potential interactors of a protein of interest can be mapped.

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Acknowledgment

This work was supported by NIH grant AR072773, a Dermatology Foundation Career Development Award grant, and a Chicago Biomedical Consortium Postdoctoral Award to B.E.P.W. and NIH grant AR062110 to S.G. Work was performed with the support of the Northwestern University Skin Disease Research Center funded by NIH grant AR057216 and Northwestern Proteomics funded by NIH grants CA060553 and GM108569.

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Authors and Affiliations

  1. Department of Dermatology, Northwestern University, Chicago, IL, USA

    Calvin J. Cable, Nihal Kaplan, Spiro Getsios & Bethany E. Perez White

  2. Department of Molecular Biosciences, Northwestern University, Chicago, IL, USA

    Paul M. Thomas

  3. Northwestern Proteomics, Northwestern University, Chicago, IL, USA

    Paul M. Thomas

  4. Skin Disease Research Center, Northwestern University, Chicago, IL, USA

    Bethany E. Perez White

Authors
  1. Calvin J. Cable
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  2. Nihal Kaplan
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  3. Spiro Getsios
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  4. Paul M. Thomas
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  5. Bethany E. Perez White
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Corresponding author

Correspondence to Bethany E. Perez White .

Editor information

Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

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© 2019 Springer Science+Business Media New York

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Cable, C.J., Kaplan, N., Getsios, S., Thomas, P.M., Perez White, B.E. (2019). Biotin Identification Proteomics in Three-Dimensional Organotypic Human Skin Cultures. In: Turksen, K. (eds) Epidermal Cells. Methods in Molecular Biology, vol 2109. Humana, New York, NY. https://doi.org/10.1007/7651_2019_239

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  • DOI: https://doi.org/10.1007/7651_2019_239

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0250-8

  • Online ISBN: 978-1-0716-0251-5

  • eBook Packages: Springer Protocols

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