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

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Abstract

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 α-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 α-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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Authors and Affiliations

  1. Photobiology Laboratory, Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, 660036, Russia

    Elena V. Eremeeva, Ludmila P. Burakova, Vasilisa V. Krasitskaya, Alexander N. Kudryavtsev, Osamu Shimomura & Ludmila A. Frank

  2. Laboratory of Bioluminescence Biotechnology, Institute of Fundamental Biology and Biotechnology, Siberian Federal University, Krasnoyarsk, 660041, Russia

    Elena V. Eremeeva, Ludmila P. Burakova, Vasilisa V. Krasitskaya, Alexander N. Kudryavtsev & Ludmila A. Frank

  3. Marine Biological Laboratory, Woods Hole, MA, 02543, USA

    Osamu Shimomura

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  1. Elena V. Eremeeva
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  2. Ludmila P. Burakova
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  3. Vasilisa V. Krasitskaya
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  6. Ludmila A. Frank
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Correspondence to Elena V. Eremeeva.

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Eremeeva, E.V., Burakova, L.P., Krasitskaya, V.V. et al. Hydrogen-bond networks between the C-terminus and Arg from the first α-helix stabilize photoprotein molecules. Photochem Photobiol Sci 13, 541–547 (2014). https://doi.org/10.1039/c3pp50369k

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  • DOI: https://doi.org/10.1039/c3pp50369k

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