Hydrogen-bond networks between the C-terminus and Arg from the first alpha-helix stabilize photoprotein molecules

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, Aequorin, Arginine, Crystallization, Escherichia coli, Hydrogen Bonding, Imidazoles, Kinetics, Luminescent Measurements, Luminescent Proteins, Mutation, Proline, Protein Stability, 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 active 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.

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Издание

Журнал: PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES

Выпуск журнала: Vol. 13, Is. 3

Номера страниц: 541-547

ISSN журнала: 1474905X

Место издания: CAMBRIDGE

Издатель: ROYAL SOC CHEMISTRY

Авторы

  • Eremeeva Elena V. (Russian Acad Sci, Inst Biophys, Siberian Branch, Photobiol Lab, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)
  • Burakova Ludmila P. (Russian Acad Sci, Inst Biophys, Siberian Branch, Photobiol Lab, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)
  • Krasitskaya Vasilisa V. (Russian Acad Sci, Inst Biophys, Siberian Branch, Photobiol Lab, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)
  • Kudryavtsev Alexander N. (Russian Acad Sci, Inst Biophys, Siberian Branch, Photobiol Lab, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)
  • Shimomura Osamu (Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia; Marine Biol Lab, Woods Hole, MA 02543 USA)
  • Frank Ludmila A. (Russian Acad Sci, Inst Biophys, Siberian Branch, Photobiol Lab, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Krasnoyarsk 660041, Russia)

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