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A Versatile Toolbox for Multiplexed Protein Micropatterning by Laser Lithography

Authors

  • Mihaela Gropeanu,

    1. Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany, Tel: +49 6131 379 516, Fax: +49 6131 379 271
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  • Maniraj Bhagawati,

    1. Universität Osnabrück, Barbarastraße 11, 49076 Osnabrück, Germany
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  • Radu A. Gropeanu,

    1. Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany, Tel: +49 6131 379 516, Fax: +49 6131 379 271
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  • Gemma M. Rodríguez Muñiz,

    1. Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany, Tel: +49 6131 379 516, Fax: +49 6131 379 271
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  • Subramanian Sundaram,

    1. Max Planck Institute für Metallforschung, Heisenbergstrasse 3, 70569 Stuttgart, Germany
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  • Jacob Piehler,

    Corresponding author
    1. Universität Osnabrück, Barbarastraße 11, 49076 Osnabrück, Germany
    • Jacob Piehler, Universität Osnabrück, Barbarastraße 11, 49076 Osnabrück, Germany.

      Aránzazu del Campo, Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany, Tel: +49 6131 379 516, Fax: +49 6131 379 271

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  • Aránzazu del Campo

    Corresponding author
    1. Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany, Tel: +49 6131 379 516, Fax: +49 6131 379 271
    • Jacob Piehler, Universität Osnabrück, Barbarastraße 11, 49076 Osnabrück, Germany.

      Aránzazu del Campo, Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany, Tel: +49 6131 379 516, Fax: +49 6131 379 271

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Abstract

Photocleavable oligohistidine peptides (POHP) allow in situ spatial organization of multiple His-tagged proteins onto surfaces functionalized with tris(nitrilotriacetic acid) (tris-NTA). Here, a second generation of POHPs is presented with improved photoresponse and site-specific covalent coupling is introduced for generating stable protein assemblies. POHPs with different numbers of histidine residues and a photocleavable linker based on the 4,5-dimethoxy-o-nitrophenyl ethyl chromophore are prepared. These peptides show better photosensitivity than the previously used o-nitrophenyl ethyl derivative. Efficient and stable caging of tris-NTA-functionalized surfaces by POHPs comprising 12 histidine residues is demonstrated by multiparameter solid-phase detection techniques. Laser lithographic uncaging by photofragmentation of the POHPs is possible with substantially reduced photodamage as compared to previous approaches. Thus, in situ micropatterning of His-tagged proteins under physiological conditions is demonstrated for the first time. In combination with a short peptide tag for a site-specific enzymatic coupling reaction, covalent immobilization of multiple proteins into target micropatterns is possible under physiological conditions.

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