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Caveolae pp 89-103 | Cite as

Biotin Proximity Labeling to Identify Protein–Protein Interactions for Cavin1

  • Carolina Mendoza-TopazEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2169)

Abstract

Biotin proximity labeling or BioID is a technique used to detect neighboring proteins, including transient and low-affinity interactions in their natural cellular environment. Here I describe the use of BioID in HeLa cells to identify proteins that can potentially interact with cavin1, one of the main components of caveolae. Briefly, the method consists in the transfection of the cells with the fusion constructs containing the promiscuous biotin ligase and cavin1 or control proteins, followed by biotin, cell lysis, affinity isolation of biotinylated proteins, biotin pull-down, and identification of the biotinylated proteins using mass spectrometry.

Key words

Caveolae Cavin1 caveolin1 Biotin proximity labeling BioID HeLa cells Mass spectrometry Biotin pull-down Protein–protein interaction 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.MRC Laboratory of Molecular BiologyCambridgeUK

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