Biotin identification proteomics in three-dimensional organotypic human skin cultures

Calvin J. Cable, Nihal Kaplan, Spiro Getsios, Paul M. Thomas, Bethany E. Perez White*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages185-197
Number of pages13
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2109
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • 3D skin culture
  • BioID
  • Biotin-identification proteomics
  • Epidermis
  • Keratinocytes
  • Protein-protein interaction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Cable, C. J., Kaplan, N., Getsios, S., Thomas, P. M., & Perez White, B. E. (2020). Biotin identification proteomics in three-dimensional organotypic human skin cultures. In Methods in Molecular Biology (pp. 185-197). (Methods in Molecular Biology; Vol. 2109). Humana Press Inc.. https://doi.org/10.1007/7651_2019_239