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Native protein nanolithography that can write, read and erase

Nature Nanotechnology volume 2, pages 220–225 (2007)Cite this article

Abstract

The development of systematic approaches to explore protein–protein interactions and dynamic protein networks is at the forefront of biological sciences. Nanopatterned protein arrays offer significant advantages for sensing applications, including short diffusion times, parallel detection of multiple targets and the requirement for only tiny amounts of sample1,2,3. Atomic force microscopy (AFM) based techniques have successfully demonstrated patterning of molecules, including stable proteins, with submicrometre resolution4,5,6,7,8,9,10,11,12,13,14,15. Here, we introduce native protein nanolithography for the nanostructured assembly of even fragile proteins or multiprotein complexes under native conditions. Immobilized proteins are detached by a novel vibrational AFM mode (contact oscillation mode) and replaced by other proteins, which are selectively self-assembled from the bulk. This nanolithography permits rapid writing, reading and erasing of protein arrays in a versatile manner. Functional protein complexes may be assembled with uniform orientation at dimensions down to 50 nm. Such fabrication of two-dimensionally arranged nano-objects with biological activity will prove powerful for proteome-wide interaction screens and single molecule/virus/cell analyses.

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Figure 1: Fabrication of rewritable protein nanoarrays on SAMs by native protein nanolithography.
Figure 2: Physical principle of COM.
Figure 3: Reversible in situ patterning of self-assembled protein monolayers.
Figure 4: Multiplexed lateral organization of protein assemblies on rewritable biochips.
Figure 5: Bioactive protein nanoarrays fabricated down to 50 nm.

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Acknowledgements

We thank Gerhard Spatz-Kümbel for technical assistance, Suman Lata and Annett Reichel for providing OG488MBP-His10, Eva Jaks for site-specifically biotinylated IFNα2, and Alart Mulder and Katrin Schulze for stimulating discussions. The work was supported by grants from the Federal Ministry of Education and Research (BMBF) (grant program: Nanobiotechnology) and the Deutsche Forschungsgemeinschaft (DFG).

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Authors and Affiliations

  1. Institute of Biochemistry, Biocenter, Johann Wolfgang Goethe-University, Max-von-Laue-Str. 9, Frankfurt, D-60438, Germany

    Ali Tinazli, Jacob Piehler & Robert Tampé

  2. Max-Planck-Institute of Biochemistry, Molecular Structural Biology, Am Klopferspitz 18, Martinsried, D-82152, Germany

    Mirjam Beuttler & Reinhard Guckenberger

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  1. Ali Tinazli
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Contributions

A.T. and R.T. conceived and designed the experiments, A.T. and M.B. performed the experiments, A.T., J.P., M.B., R.G., and R.T. analysed the data, and A.T. and R.T. co-wrote the paper.

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Correspondence to Robert Tampé.

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Tinazli, A., Piehler, J., Beuttler, M. et al. Native protein nanolithography that can write, read and erase. Nature Nanotech 2, 220–225 (2007). https://doi.org/10.1038/nnano.2007.63

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