caa a22 4500 378893130 CHVBK 20180305123500.0 cr unu---uuuuu 161128e20040205xx s 000 0 eng 10.1515/BC.2004.038 doi (NATIONALLICENCE)gruyter-10.1515/BC.2004.038 Probing the role of tryptophans in Aequorea victoria green fluorescent proteins with an expanded genetic code [Elektronische Daten] [N. Budisa, P. P. Pal, S. Alefelder, P. Birle, T. Krywcun, M. Rubini, W. Wenger, J. H. Bae, T. Steiner] 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. Copyright © 2004 by Walter de Gruyter GmbH & Co. KG Biochemistry nationallicence Molecular biology nationallicence Cellular biology nationallicence Budisa N. aut Pal P. P. aut Alefelder S. aut Birle P. aut Krywcun T. aut Rubini M. aut Wenger W. aut Bae J. H. aut Steiner T. aut Biological Chemistry Walter de Gruyter 385/2(2004-02-05), 191-202 1431-6730 385:2<191 2004 385 bchm https://doi.org/10.1515/BC.2004.038 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/BC.2004.038 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Budisa N. aut NATIONALLICENCE 700 1- Pal P. P. aut NATIONALLICENCE 700 1- Alefelder S. aut NATIONALLICENCE 700 1- Birle P. aut NATIONALLICENCE 700 1- Krywcun T. aut NATIONALLICENCE 700 1- Rubini M. aut NATIONALLICENCE 700 1- Wenger W. aut NATIONALLICENCE 700 1- Bae J. H. aut NATIONALLICENCE 700 1- Steiner T. aut NATIONALLICENCE 773 0- Biological Chemistry Walter de Gruyter 385/2(2004-02-05), 191-202 1431-6730 385:2<191 2004 385 bchm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter