التفاصيل البيبلوغرافية
| العنوان: |
A Green Fluorescent Protein Containing a QFG Tri-Peptide Chromophore: Optical Properties and X-Ray Crystal Structure. |
| المؤلفون: |
Battad, Jion M., Traore, Daouda A. K., Byres, Emma, Rossjohn, Jamie, Devenish, Rodney J., Olsen, Seth, Wilce, Matthew C. J., Prescott, Mark |
| المصدر: |
PLoS ONE; Oct2012, Vol. 7 Issue 10, Special section p1-14, 14p |
| مصطلحات موضوعية: |
PROTEIN research, TRIPEPTIDES, TYROSINE, PHENYLALANINE, FLUORESCENT proteins, CHROMOPHORES |
| مستخلص: |
Rtms5 is an deep blue weakly fluorescent GFP-like protein (λAbsmax, 592 nm; λEmmax, 630nm; ΦF, 0.004) that contains a 66Gln-Tyr-Gly chromophore tripeptide sequence. We investigated the optical properties and structure of two variants, Rtms5Y67F and and Rtms5Y67F/H146S in which the tyrosine at position 67 was substituted by a phenylalanine. Compared to the parent proteins the optical spectra for these new variants were significantly blue-shifted. Rtms5Y67F spectra were characterised by two absorbing species (λAbsmax, 440 nm and 513 nm) and green fluorescence emission (λExmax, 440 nm; λEmmax, 508 nm; ΦF, 0.11), whilst Rtms5Y67F/H146S spectra were characterised by a single absorbing species (λAbsmax, 440 nm) and a relatively high fluorescence quantum yield (ΦF, 0.75; λExmax, 440 nm; λEmmax, 508 nm). The fluorescence emissions of each variant were remarkably stable over a wide range of pH (3-11). These are the first GFP-like proteins with green emissions (500-520 nm) that do not have a tyrosine at position 67. The X-ray crystal structure of each protein was determined to 2.2 Å resolution and showed that the benzylidine ring of the chromophore, similar to the 4-hydroxybenzylidine ring of the Rtms5 parent, is non-coplanar and in the trans conformation. The results of chemical quantum calculations together with the structural data suggested that the 513 nm absorbing species in Rtms5Y67F results from an unusual form of the chromophore protonated at the acylimine oxygen. These are the first X-ray crystal structures for fluorescent proteins with a functional chromophore containing a phenylalanine at position 67. [ABSTRACT FROM AUTHOR] |
|
Copyright of PLoS ONE is the property of Public Library of Science and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) |
| قاعدة البيانات: |
Complementary Index |
Full Text Finder