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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Sies, Helmut / Turk, Boris / Wittinghofer, Alfred


SCImago Journal Rank (SJR) 2015: 1.607
Source Normalized Impact per Paper (SNIP) 2015: 0.751
Impact per Publication (IPP) 2015: 2.609

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1437-4315
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Probing the role of tryptophans in Aequorea victoria green fluorescent proteins with an expanded genetic code

N. Budisa / P. P. Pal / S. Alefelder / P. Birle / T. Krywcun / M. Rubini / W. Wenger / J. H. Bae / T. Steiner

Citation Information: Biological Chemistry. Volume 385, Issue 2, Pages 191–202, ISSN (Print) 1431-6730, DOI: 10.1515/BC.2004.038, June 2005

Publication History

Published Online:
2005-06-01

Abstract

The expanded genetic code in combination with sitedirected mutagenesis was used to probe spectroscopic and structural roles of tryptophan (Trp) residues in Aequorea victoria green fluorescent proteins (avGFPs). Nine different halogen-, chalcogen-, and methyl-containing Trp isosteric analogues and surrogates were incorporated into avGFPs containing indole moieties in, and outside of, the chromophore, by the use of the selective pressure incorporation method. Such isosteric replacements introduced minimal local geometry changes in indole moieties, often to the level of single atomic exchange (atomic mutation) and do not affect three-dimensional structures of avGFPs but induce changes in spectral properties. Our approach offers a new platform to re-evaluate issues like resonance transfer, mechanisms of chromophore formation and maturation, as well as the importance of local geometry and weak sulphuraromatic interactions for avGFP spectral properties and structural stability. The library of novel tailor-made avGFP mutants and variants generated in this work has demonstrated not only the potentials of the expanded genetic code to study spectroscopic functions, but also a new approach to generate tailor-made proteins with interesting and useful spectral properties.

Citing Articles

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[12]
S. Lepthien, M. G. Hoesl, L. Merkel, and N. Budisa
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[13]
Sandra Lepthien, Birgit Wiltschi, Bojana Bolic, and Nediljko Budisa
Applied Microbiology and Biotechnology, 2006, Volume 73, Number 4, Page 740
[14]
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