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Quantifying Biomolecular Recognition with Site-Specific 2D Infrared Probes


Johnson, Philip J M; Koziol, Klemens L; Hamm, Peter (2017). Quantifying Biomolecular Recognition with Site-Specific 2D Infrared Probes. Journal of Physical Chemistry Letters, 8(10):2280-2284.

Abstract

Azidohomoalanine (Aha) is an unnatural amino acid containing an infrared active azido side chain group that can, through frequency shifts of the azido stretch vibration, act as a probe of local structure. To realize the potential of such structural probes for protein science, we have developed a two-dimensional infrared spectrometer employing fast mechanical scanning and intrinsic phasing of the resulting spectra, leading to a lower sensitivity limit of similar to 100 mu OD level samples. Using this approach, we quantify the biomolecular recognition between a PDZ2 domain and two Aha-mutated peptides. It is shown that this method can distinguish different binding modes and that the energetics of binding can be determined.

Abstract

Azidohomoalanine (Aha) is an unnatural amino acid containing an infrared active azido side chain group that can, through frequency shifts of the azido stretch vibration, act as a probe of local structure. To realize the potential of such structural probes for protein science, we have developed a two-dimensional infrared spectrometer employing fast mechanical scanning and intrinsic phasing of the resulting spectra, leading to a lower sensitivity limit of similar to 100 mu OD level samples. Using this approach, we quantify the biomolecular recognition between a PDZ2 domain and two Aha-mutated peptides. It is shown that this method can distinguish different binding modes and that the energetics of binding can be determined.

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Additional indexing

Item Type:Journal Article, refereed, further contribution
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Language:English
Date:18 May 2017
Deposited On:09 Mar 2018 12:59
Last Modified:04 May 2018 00:00
Publisher:American Chemical Society (ACS)
ISSN:1948-7185
Funders:European Research Council (ERC) Advanced Investigator Grant (DYNALLO), Swiss National Science Foundation (SNF) through the NCCR MUST
OA Status:Green
Publisher DOI:https://doi.org/10.1021/acs.jpclett.7b00742
Project Information:
  • : Funder
  • : Grant ID
  • : Project TitleEuropean Research Council (ERC) Advanced Investigator Grant (DYNALLO)
  • : FunderSNSF
  • : Grant ID
  • : Project TitleSwiss National Science Foundation (SNF) through the NCCR MUST

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