Тип публикации: статья из журнала
Год издания: 2014
Идентификатор DOI: 10.1039/c3pp50369k
Ключевые слова: aequorin, arginine, coelenterazine, imidazole derivative, obelin, photoprotein, proline, pyrazine derivative, recombinant protein, article, chemistry, crystallization, Escherichia coli, genetics, hydrogen bond, kinetics, luminescence, mutation, protein secondary structure, protein stability, Hydrogen Bonding, Imidazoles, Luminescent Measurements, Luminescent Proteins, Protein Structure, Secondary, Pyrazines, Recombinant Proteins
Аннотация: Previous studies have stated that aequorin loses most of its bioluminescence activity upon modification of the C-terminus, thus limiting the production of photoprotein fusion proteins at its N-terminus. In the present work, we investigate the importance of the C-terminal proline and the hydrogen bonds it forms for photoprotein actiПоказать полностьюve complex formation, stability and functional activity. According to the crystal structures of obelin and aequorin, two Ca2+-regulated photoproteins, the carboxyl group of the C-terminal Pro forms two hydrogen bonds with the side chain of Arg21 (Arg15 in aequorin case) situated in the first a-helix. Whereas, deletion or substitution of the C-terminal proline could noticeably change the bioluminescence activity, stability or the yield of an active photoprotein complex. Therefore, modifications of the first alpha-helix Arg has a clear destructive effect on the main photoprotein properties. A C-terminal hydrogen-bond network is proposed to be important for the stability of photoprotein molecules towards external disturbances, when taking part in the formation of locked protein conformations and isolation of coelenterazine-binding cavities.
Журнал: PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES
Выпуск журнала: Vol. 13, Is. 3
Номера страниц: 541-547
ISSN журнала: 1474905X
Место издания: CAMBRIDGE
Издатель: ROYAL SOC CHEMISTRY