Probing cellular protein complexes using single-molecule pull-down

Ankur Jain, Ruijie Liu, Biswarathan Ramani, Edwin Arauz, Yuji Ishitsuka, Kaushik Ragunathan, Jeehae Park, Jie Chen, Yang K. Xiang () and Taekjip Ha ()
Additional contact information
Ankur Jain: Center for Biophysics and Computational Biology and Institute for Genomic Biology, University of Illinois at Urbana-Champaign
Ruijie Liu: University of Illinois at Urbana-Champaign
Biswarathan Ramani: University of Illinois at Urbana-Champaign
Edwin Arauz: University of Illinois at Urbana-Champaign
Yuji Ishitsuka: University of Illinois at Urbana-Champaign
Kaushik Ragunathan: Center for Biophysics and Computational Biology and Institute for Genomic Biology, University of Illinois at Urbana-Champaign
Jeehae Park: Center for Biophysics and Computational Biology and Institute for Genomic Biology, University of Illinois at Urbana-Champaign
Jie Chen: University of Illinois at Urbana-Champaign
Yang K. Xiang: University of Illinois at Urbana-Champaign
Taekjip Ha: Center for Biophysics and Computational Biology and Institute for Genomic Biology, University of Illinois at Urbana-Champaign

Nature, 2011, vol. 473, issue 7348, 484-488

Abstract: Abstract Proteins perform most cellular functions in macromolecular complexes. The same protein often participates in different complexes to exhibit diverse functionality. Current ensemble approaches of identifying cellular protein interactions cannot reveal physiological permutations of these interactions. Here we describe a single-molecule pull-down (SiMPull) assay that combines the principles of a conventional pull-down assay with single-molecule fluorescence microscopy and enables direct visualization of individual cellular protein complexes. SiMPull can reveal how many proteins and of which kinds are present in the in vivo complex, as we show using protein kinase A. We then demonstrate a wide applicability to various signalling proteins found in the cytosol, membrane and cellular organelles, and to endogenous protein complexes from animal tissue extracts. The pulled-down proteins are functional and are used, without further processing, for single-molecule biochemical studies. SiMPull should provide a rapid, sensitive and robust platform for analysing protein assemblies in biological pathways.

Date: 2011
References: Add references at CitEc
Citations: View citations in EconPapers (2)

Downloads: (external link)
https://www.nature.com/articles/nature10016 Abstract (text/html)
Access to the full text of the articles in this series is restricted.

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:nature:v:473:y:2011:i:7348:d:10.1038_nature10016

Ordering information: This journal article can be ordered from
https://www.nature.com/

DOI: 10.1038/nature10016

Access Statistics for this article

Nature is currently edited by Magdalena Skipper

More articles in Nature from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().