Single-molecule pull-down (SiMPull) for new-age biochemistry: Methodology and biochemical applications of single-molecule pull-down (SiMPull) for probing biomolecular interactions in crude cell extracts

Vasudha Aggarwal, Taekjip Ha

Research output: Contribution to journalArticlepeer-review

Abstract

Macromolecular interactions play a central role in many biological processes. Protein-protein interactions have mostly been studied by co-immunoprecipitation, which cannot provide quantitative information on all possible molecular connections present in the complex. We will review a new approach that allows cellular proteins and biomolecular complexes to be studied in real-time at the single-molecule level. This technique is called single-molecule pull-down (SiMPull), because it integrates principles of conventional immunoprecipitation with the powerful single-molecule fluorescence microscopy. SiMPull is used to count how many of each protein is present in the physiological complexes found in cytosol and membranes. Concurrently, it serves as a single-molecule biochemical tool to perform functional studies on the pulled-down proteins. In this review, we will focus on the detailed methodology of SiMPull, its salient features and a wide range of biological applications in comparison with other biosensing tools.

Original languageEnglish (US)
Pages (from-to)1109-1119
Number of pages11
JournalBioEssays
Volume36
Issue number11
DOIs
StatePublished - Nov 1 2014

Keywords

  • Co-immunoprecipitation
  • Fluorescence
  • Protein complex
  • Pull-down
  • Single molecule
  • Western blotting

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

Fingerprint

Dive into the research topics of 'Single-molecule pull-down (SiMPull) for new-age biochemistry: Methodology and biochemical applications of single-molecule pull-down (SiMPull) for probing biomolecular interactions in crude cell extracts'. Together they form a unique fingerprint.

Cite this