A TurboID-based proximity labelling approach for identifying the DNA-binding proteins

STAR Protoc. 2023 Mar 17;4(1):102139. doi: 10.1016/j.xpro.2023.102139. Epub 2023 Feb 26.

Abstract

Biotin proximity labeling is a technique based on the TurboID enzyme that can be used to capture weak or dynamic interactions that had previously not been used to map proteins interacting with a specific DNA sequence. Here, we present a protocol for identifying specific DNA-sequence-binding proteins. We describe steps for biotin labeling of DNA-binding proteins, protein enrichment and sodium dodecyl sulfate polyacrylamide gel electrophoresis separation, and proteomic analysis. For complete details on the use and execution of this protocol, please refer to Wei et al. (2022).1.

Keywords: Mass Spectrometry; Molecular/Chemical Probes; Protein Biochemistry; Proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin*
  • DNA-Binding Proteins*
  • Proteomics / methods

Substances

  • Biotin
  • DNA-Binding Proteins