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 language | English (US) |
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Pages (from-to) | 1109-1119 |
Number of pages | 11 |
Journal | BioEssays |
Volume | 36 |
Issue number | 11 |
DOIs | |
State | Published - 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