Abstract
The non-catalytic COOH-terminal module formed after proteolytic cleavage of full-length mammalian tyrosyl-tRNA synthetase displays dual function: tRNA binding ability and cytokine activity. With the aim to explore the intramolecular dynamics of C-module in solution we used fluorescence spectroscopy to study conformational changes of isolated protein. We used information from fluorescence spectra and computational model for characterization of a microenvironment of a single tryptophan residue (Trp144). Its fluorescence parameters and protection from quenching by Cs+ ions indicate the internal localization—buried into protein globule. The fluorescence quenching of Trp144 by acrylamide suggests rapid conformation dynamics of the C-module in nanosecond time scale. The temperature-induced conformational changes in the C-module were monitored by the fluorescence measurements of Trp144 emission and by red-edge excitation shift. An emission maximum shift up to ∼349 nm and significant decrease of the red-edge shift effect at 37–52 °C indicated a major conformational transition of Trp144 from buried native state into highly relaxing polar solvent environment.
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Acknowledements
We are grateful to Professors A.P. Demchenko and Y. Mely, Dr. E. Domanov for helpful discussion; Dr. A. Dubrovsky for pET30a-YCD3 plasmid preparation: K. Odynets for building tha cm5 model. This work was supported by grant No 5.07/200 from the Ministry of Science and Education of Ukraine.
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Kordysh, M., Kornelyuk, A. Conformational Flexibility of Cytokine-Like C-Module of Tyrosyl-tRNA Synthetase Monitored by Trp144 Intrinsic Fluorescence. J Fluoresc 16, 705–711 (2006). https://doi.org/10.1007/s10895-006-0113-9
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DOI: https://doi.org/10.1007/s10895-006-0113-9