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Efficient Isotope E...
Efficient Isotope Editing of Proteins for Site-Directed Vibrational Spectroscopy
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- Peuker, Sebastian (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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- Andersson, Hanna, 1979 (author)
- Uppsala universitet,Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,Organisk kemi
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- Gustavsson, Emil, 1987 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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- Maiti, K. S. (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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- Kania, R (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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- Karim, Alavi, 1986 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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- Niebling, Stephan (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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- Pedersen, Anders, 1976 (author)
- Gothenburg University,Göteborgs universitet,Svenskt NMR-centrum vid Göteborgs universitet,Swedish NMR Centre at Göteborg University,Swedish NMR Centre at the University of Gothenburg, Gothenburg, Sweden
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- Erdelyi, Mate, 1975 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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- Westenhoff, Sebastian, 1978 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology,University of Gothenburg, Gothenburg, Sweden
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(creator_code:org_t)
- 2016-02-10
- 2016
- English.
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In: Journal of the American Chemical Society. - : American Chemical Society (ACS). - 0002-7863 .- 1520-5126. ; 138:7, s. 2312-2318
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https://doi.org/10.1...
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Abstract
Subject headings
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- Vibrational spectra contain unique information on protein structure and dynamics. However, this information is often obscured by spectral congestion, and site-selective information is not available. In principle, sites of interest can be spectrally identified by isotope shifts, but site-specific isotope labeling of proteins is today possible only for favorable amino acids or with prohibitively low yields. Here we present an efficient cell-free expression system for the site-specific incorporation of any isotope-labeled amino acid into proteins. We synthesized 1.6 mg of green fluorescent protein with an isotope-labeled tyrosine from 100 mL of cell free reaction extract. We unambiguously identified spectral features of the tyrosine in the fingerprint region of the time-resolved infrared absorption spectra. Kinetic analysis confirmed the existence of an intermediate state between photoexcitation and proton transfer that lives for 3 ps. Our method lifts vibrational spectroscopy of proteins to a higher level of structural specificity.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- NATURVETENSKAP -- Kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences (hsv//eng)
- NATURVETENSKAP -- Kemi -- Organisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Organic Chemistry (hsv//eng)
Keyword
- green fluorescent protein
- 2-dimensional infrared-spectroscopy
- unnatural amino-acids
- proton-transfer
- chemical aminoacylation
- transfer-rnas
- genetic-code
- active-site
- dynamics
- expression
- Chemistry
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Peuker, Sebastia ...
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Andersson, Hanna ...
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Gustavsson, Emil ...
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Maiti, K. S.
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Kania, R
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Karim, Alavi, 19 ...
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show more...
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Niebling, Stepha ...
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Pedersen, Anders ...
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Erdelyi, Mate, 1 ...
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Westenhoff, Seba ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Chemical Science ...
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Chemical Science ...
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and Organic Chemistr ...
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Journal of the A ...
- By the university
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University of Gothenburg
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Uppsala University