Abstract
The L232H mutant of the enhanced green fluorescent protein (EGFP) was expressed and crystallized. An X-ray diffraction data set was collected from the crystals to 1.53 Å resolution. An analysis of the three-dimensional structure revealed a stacking interaction between the amino-acid residues Н78 and Н232, which contributes to the fastening of the C-terminal region of the protein in the vicinity of the chromophore and influences chromophore maturation of hybrid fluorescent proteins produced by fusion of the target proteins with the C-terminus of EGFP. This hypothesis was experimentally confirmed by investigating chromophore maturation of the hybrid proteins fused to the N- and C-termini of EGFP and EGFP-L232H.
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References
R. N. Day and M. W. Davidson, Chem. Soc. Rev. 38, 4 (2009).
G. Zhang, V. Gurtu, and S. R. Kain, Biochem. Biophys. Res. Commun. 227, 707 (1996).
D. C. Prasher, V. K. Eckenrode, W. W. Ward, et al., Gene 111, 229 (1992).
B. P. Cormack, R. H. Valdivia, and S. Falkow, Gene 173, 33 (1996).
M. R. Green and J. Sambrook, Molecular Cloning: A Laboratory Manual (Fourth Edition) (Cold Spring Harbor Laboratory Press, 2012).
V. Timofeev, E. Slutskaya, M. Gorbacheva, et al., Acta Crystallogr. F 71, 951 (2015).
A. Yu. Nikolaeva, V. I. Timofeev, K. M. Boiko, et al., Crystallogr. Rep. 60, 880 (2015).
D. A. Altukhov, Yu. K. Agapova, A. V. Vlaskina, et al., Vestn. Mosk. Univ. 57 (4), 226 (2016).
D. A. Altukhov, A. A. Talyzina, Y. K. Agapova, et al., J. Biomol. Struct. Dyn. (2016); doi 10.1080/07391102.2016.1264893
U. K. Laemmli, Nature 227, 680 (1970).
L. Syrovy and Z. J. Hodny, Chromatography 569, 175 (1991).
M. Ormö, A. B. Cubitt, K. Kallio, et al., Science 273, 1392 (1996).
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Original Russian Text © A.A. Simanovskaya, T.V. Ivashina, Y.S. Zeifman, T.V. Fateeva, M.V. Krukova, A.N. Popov, G.S. Kachalova, T.V. Rakitina, 2018, published in Kristallografiya, 2018, Vol. 63, No. 2, pp. 208–212.
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Simanovskaya, A.A., Ivashina, T.V., Zeifman, Y.S. et al. Mutation L232H Promotes Chromophore Maturation of EGFP-Based Fluorescent Fusion Proteins. Crystallogr. Rep. 63, 191–195 (2018). https://doi.org/10.1134/S1063774518020256
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DOI: https://doi.org/10.1134/S1063774518020256