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Proceedings Paper

Nanoscale fabrication of a peptide layer using an AFM probe
Author(s): Chikashi Nakamura; Chie Miyamoto; Ikuo Obataya; Noriyuki Nakamura; Jun Miyake
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Paper Abstract

Scanning probe microscopy has been applied in many studies to manipulate atoms or molecules. In particular, force spectroscopy using an atomic force microscope (AFM) is a powerful tool to elucidate intermolecular or intramolecular interactions and provide mechanical information. If enzymes could retain their activity when immobilized on probes, not only could enzyme-substrate interactions be investigated but also the probes could be used for precise biomolecular manipulation at the nano-scale. In our study, a method based on "Enzymatic Nanolithography" was successfully performed in a buffered solution using Staphylococcal serine V8 protease and AFM. To estimate the fabricating activity of the protease immobilized on the AFM tip to peptides immobilized on a substrate, we designed and synthesized peptides that showed enzymatic action specific to the protease. When the protease digested the reporter peptide a quencher residue was released from the main flame of the peptide and resulted in fluorescence. In the designed 9 mer peptides, TAMRA functioned as a good quencher for FAM. After contact of the protease-immobilized tip to the reporter peptide layer, a fluorescent area was observed by microscopic imaging.

Paper Details

Date Published: 29 December 2004
PDF: 7 pages
Proc. SPIE 5593, Nanosensing: Materials and Devices, (29 December 2004); doi: 10.1117/12.578115
Show Author Affiliations
Chikashi Nakamura, National Institute of Advanced Industrial Science and Technology (Japan)
Tokyo Univ. of Agriculture and Technology (Japan)
Chie Miyamoto, National Institute of Advanced Industrial Science and Technology (Japan)
Tokyo Univ. of Agriculture and Technology (Japan)
Ikuo Obataya, National Institute of Advanced Industrial Science and Technology (Japan)
Noriyuki Nakamura, National Institute of Advanced Industrial Science and Technology (Japan)
Tokyo Univ. of Agriculture and Technology (Japan)
Jun Miyake, National Institute of Advanced Industrial Science and Technology (Japan)
Tokyo Univ. of Agriculture and Technology (Japan)


Published in SPIE Proceedings Vol. 5593:
Nanosensing: Materials and Devices
M. Saif Islam; Achyut K. Dutta, Editor(s)

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