Mutation L232H Promotes Chromophore Maturation of EGFP-Based Fluorescent Fusion Proteins

Journal Article · Thu Mar 15 04:00:00 EDT 2018 · Crystallography Reports

DOI:https://doi.org/10.1134/S1063774518020256· OSTI ID:22758264

Simanovskaya, A. A. ^[1]; Ivashina, T. V. ^[2]; Zeifman, Y. S.; Fateeva, T. V.; Krukova, M. V. ^[1]; Popov, A. N. ^[3]; Kachalova, G. S.

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1. National Research Centre “Kurchatov Institute,” (Russian Federation)
2. Russian Academy of Sciences, Skryabin Institute of Biochemistry and Physiology of Microorganisms (Russian Federation)
3. European Synchrotron Radiation Facility (France)

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.

• OSTI ID: 22758264
• Journal Information: Crystallography Reports, Journal Name: Crystallography Reports Journal Issue: 2 Vol. 63; ISSN CYSTE3; ISSN 1063-7745
• Country of Publication: United States
• Language: English

Related Subjects

75 CONDENSED MATTER PHYSICS
SUPERCONDUCTIVITY AND SUPERFLUIDITY
AMINO ACIDS
CRYSTALS
FASTENING
FLUORESCENCE
HYPOTHESIS
MATURATION
MUTATIONS
PROTEINS
RESOLUTION
THREE-DIMENSIONAL LATTICES
X-RAY DIFFRACTION