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Devising self-assembled-monolayers for surface-enhanced infrared spectroscopy of pH-driven poly-L-lysine conformational changes

MPG-Autoren
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Stanglmair,  Christoph
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;

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Pacholski,  Claudia
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;
Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany;

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Zitation

Fallah, M. A., Stanglmair, C., Pacholski, C., & Hauser, K. (2016). Devising self-assembled-monolayers for surface-enhanced infrared spectroscopy of pH-driven poly-L-lysine conformational changes. Langmuir, 32(29), 7356-7364. doi:10.1021/acs.langmuir.6b01742.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-002C-46A7-F
Zusammenfassung
Surface-enhanced infrared absorption spectroscopy (SEIRA) is applied to study protein conformational changes. In general, the appropriate functionalization of metal surfaces with biomolecules remains a challenge if the conformation and activity of the biomolecule shall be preserved. Here we present a SEIRA study to monitor pH-induced conformational changes of poly-l-lysine (PLL) covalently bound to a thin gold layer via self-assembled monolayers (SAMs). We demonstrate that the composition of the SAM is crucial. A SAM of 11-mercaptoundecanonic acid (MUA) can link PLL to the gold layer, but pH-driven conformational transitions were hindered compared to poly-l-lysine in solution. To address this problem, we devised a variety of SAMs, i.e., mixed SAMs of MUA with either octanethiol (OT) or 11-mercapto-1-undecanol (MUoL) and furthermore SAMs of MT(PEG)4 and NHS-PEG10k-SH. These mixed SAMs modify the surface properties by changing the polarity and the morphology of the surface present to nearby PLL molecules. Our experiments reveal that mixed SAMs of MUA-MUoL and SAMs of NHS-PEG10k-SH-MT(PEG)4 are suitable to monitor pH-driven conformational changes of immobilized PLL. These SAMs might be applicable for chemoselective protein immobilization in general.