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Research article2006Peer reviewed

Direct observation of individual endogenous protein complexes in situ by proximity ligation

Soderberg O, Gullberg M, Jarvius M, Ridderstrale K, Leuchowius KJ, Jarvius J, Wester K, Hydbring P, Bahram F, Larsson LG, Landegren U

Abstract

Cellular processes can only be understood as the dynamic interplay of molecules. There is a need for techniques to monitor interactions of endogenous proteins directly in individual cells and tissues to reveal the cellular and molecular architecture and its responses to perturbations. Here we report our adaptation of the recently developed proximity ligation method to examine the subcellular localization of protein-protein interactions at single-molecule resolution. Proximity probes-oligonucleotides attached to antibodies against the two target proteins-guided the formation of circular DNA strands when bound in close proximity. The DNA circles in turn served as templates for localized rolling-circle amplification (RCA), allowing individual interacting pairs of protein molecules to be visualized and counted in human cell lines and clinical specimens. We used this method to show specific regulation of protein-protein interactions between endogenous Myc and Max oncogenic transcription factors in response to interferon-gamma (IFN-gamma) signaling and low-molecular-weight inhibitors

Published in

Nature Methods
2006, Volume: 3, number: 12, pages: 995-1000
Publisher: NATURE PUBLISHING GROUP

      SLU Authors

        • UKÄ Subject classification

        Agricultural Science
        Food Science
        Forest Science

        Publication identifier

        DOI: https://doi.org/10.1038/nmeth947

        Permanent link to this page (URI)

        https://res.slu.se/id/publ/14444