Тип публикации: статья из журнала
Год издания: 2013
Идентификатор DOI: 10.1016/j.jphotobiol.2013.08.012
Ключевые слова: Bioluminescence, Coelenterazine, Obelin, Aequorin, Photoprotein, 2 hydroperoxy adduct, amide, histidine, imidazopyrazine derivative, peroxy radical, proton, tryptophan, tyrosine, unclassified drug, amino acid substitution, article, binding affinity, chemical reaction kinetics, complex formation, controlled study, crystal structure, nonhuman, priority journal, protein structure, protein synthesis, site directed mutagenesis, Apoproteins, Calcium, Imidazoles, Kinetics, Luminescent Proteins, Models, Molecular, Mutation, Protein Binding, Protein Conformation, Pyrazines, Spectrometry, Fluorescence
Аннотация: Bioluminescence of a variety of marine organisms is caused by monomeric Ca2+-regulated photoproteins, to which a peroxy-substituted coelenterazine, 2-hydroperoxycoelenterazine, is firmly bound. From the spatial structure the side chains of Tyr138, His175, Trp179, and Tyr190 of obelin are situated within the substrate-binding pocket at hydrogen bond distances with different atoms of the 2-hydroperoxycoelenterazine. Here we characterized several obelin mutants with substitutions of these residues regarding their bioluminescence, coelenterazine binding, and kinetics of active obelin formation. We demonstrate that Tyr138, His175, Trp179, and Tyr190 are all important for coelenterazine activation; substitution of any of these residues leads to significant decrease of the apparent reaction rate. The hydrogen bond network formed by Tyr138, Trp179 and Tyr190 participates in the proper positioning of coelenterazine in the active site and subsequent stabilization of the 2-hydroperoxy adduct of coelenterazine. His175 might serve as a proton shuttle during 2-hydroperoxycoelenterazine formation. (C) 2013 Elsevier B.V. All rights reserved.
Издание
Журнал: JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY
Выпуск журнала: Vol. 127
Номера страниц: 133-139
ISSN журнала: 10111344
Место издания: LAUSANNE
Издатель: ELSEVIER SCIENCE SA
Персоны
- Eremeeva Elena V. (Russian Acad Sci, Photobiol Lab, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia; Wageningen Univ, Biochem Lab, NL-6703 HA Wageningen, Netherlands; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)
- Markova Svetlana V. (Russian Acad Sci, Photobiol Lab, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)
- van Berkel Willem J. H. (Wageningen Univ, Biochem Lab, NL-6703 HA Wageningen, Netherlands)
- Vysotski Eugene S. (Russian Acad Sci, Photobiol Lab, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)
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