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Fibrinogen anchors for micropatterning of active proteins and subcellular receptor relocalisation

Joseph L. Watson, Samya Aich, View ORCID ProfileBenjamí Oller Salvia, Andrew A. Drabek, Stephen C. Blacklow, Jason Chin, View ORCID ProfileEmmanuel Derivery
doi: https://doi.org/10.1101/2020.09.04.256875
Joseph L. Watson
1MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, UK
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Samya Aich
1MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, UK
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Benjamí Oller Salvia
1MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, UK
4Institut Químic de Sarrià, Universitat Ramon Llull, 08017 Barcelona, Spain
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  • ORCID record for Benjamí Oller Salvia
Andrew A. Drabek
2Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA
3Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA
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Stephen C. Blacklow
2Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA
3Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA
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Jason Chin
1MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, UK
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Emmanuel Derivery
1MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, UK
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  • ORCID record for Emmanuel Derivery
  • For correspondence: derivery@mrc-lmb.cam.ac.uk
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Abstract

Protein micropatterning allows proteins to be precisely deposited onto a substrate of choice, and is now routinely used in cell biology and in vitro reconstitution. However, a drawback of current technology is that micropatterning efficiency can be variable between proteins, and that proteins may lose activity on the micropatterns. Here, we describe a general method to enable micropatterning of virtually any protein at high specificity and homogeneity while maintaining its activity. Our method is based on an anchor that micropatterns well, Fibrinogen, which we functionalized to bind to common purification tags. This enhances micropatterning on various substrates, facilitates multiplexed micropatterning, and dramatically improves the on-pattern activity of fragile proteins like molecular motors. Furthermore, it enhances the micropatterning of hard to micropattern cells. Last, this method enables subcellular micropatterning, whereby complex micropatterns simultaneously control cell shape and the distribution of transmembrane receptors within that cell. Altogether, these results open new avenues for cell biology.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted September 21, 2020.
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Fibrinogen anchors for micropatterning of active proteins and subcellular receptor relocalisation
Joseph L. Watson, Samya Aich, Benjamí Oller Salvia, Andrew A. Drabek, Stephen C. Blacklow, Jason Chin, Emmanuel Derivery
bioRxiv 2020.09.04.256875; doi: https://doi.org/10.1101/2020.09.04.256875
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Fibrinogen anchors for micropatterning of active proteins and subcellular receptor relocalisation
Joseph L. Watson, Samya Aich, Benjamí Oller Salvia, Andrew A. Drabek, Stephen C. Blacklow, Jason Chin, Emmanuel Derivery
bioRxiv 2020.09.04.256875; doi: https://doi.org/10.1101/2020.09.04.256875

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