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Longer characteristic wavelength in a novel engineered photoprotein Mnemiopsin 2

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Abstract

We designed two mutants of photoprotein Mnemiopsin 2 (Mn2) including M52I and V144I, where the mutations were applied in the EF-hand loops I and III. Far-UV CD measurements demonstrated that the stability of the helices in the wild-type (WT) protein is greater compared with the mutants. Heat-induced denaturation experiments in the apo-form of photoproteins showed that WT Mn2 has higher value of the enthalpy change for the unfolding process, indicating that it has more stabilizing interaction compared with mutants. According to the activity measurement data, both mutants, particularly V144I have lower initial intensity as well as slower decay rate as compared with the WT photoprotein. Importantly, it was found that V144I variant shows 25 nm of red shift in the characteristic wavelengths as compared with the WT photoprotein. This finding can be considered as an advantage for in vivo application of photoprotein for imaging purposes. It concluded that this position on loop III of Mn2 is a hotspot point for characteristic wavelength determination. However, further research on this mutant is needed for making stable variants of Mn2 with novel optical features.

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Acknowledgements

We acknowledge the research council of the University of Zanjan.

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Authors and Affiliations

  1. Department of Biology, Faculty of Sciences, University of Zanjan, Zanjan, Iran

    Mahsa Hematyar & Vahab Jafarian

  2. Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    Akram Shirdel

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  1. Mahsa Hematyar
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  2. Vahab Jafarian
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Correspondence to Vahab Jafarian or Akram Shirdel.

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Partial financial support was received from the research council of the University of Zanjan. The authors declare they have no financial interests, and they have no relevant financial or non-financial interests to disclose.

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Hematyar, M., Jafarian, V. & Shirdel, A. Longer characteristic wavelength in a novel engineered photoprotein Mnemiopsin 2. Photochem Photobiol Sci 21, 1031–1040 (2022). https://doi.org/10.1007/s43630-022-00191-6

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