6ZQO

EYFP mutant - F165G

Summary for 6ZQO

• Entry DOI: 10.2210/pdb6zqo/pdb
• Descriptor: G protein/GFP fusion protein, SULFATE ION (3 entities in total)
• Functional Keywords: alternative routes of post-translation chemistry, fluorescent protein
• Biological source: Recombinant vesicular stomatitis Indiana virus rVSV-G/GFP
• Total number of polymer chains: 1
• Total formula weight: 28417.97

Authors

Pletnev, V.Z.,Pletnev, S.V.,Pletneva, N.V. (deposition date: 2020-07-10, release date: 2021-06-16, Last modification date: 2024-10-16)

Primary citation

Pletneva, N.V.,Maksimov, E.G.,Protasova, E.A.,Mamontova, A.V.,Simonyan, T.R.,Ziganshin, R.H.,Lukyanov, K.A.,Muslinkina, L.,Pletnev, S.,Bogdanov, A.M.,Pletnev, V.Z.
Amino acid residue at the 165th position tunes EYFP chromophore maturation. A structure-based design.
Comput Struct Biotechnol J, 19:2950-2959, 2021

PubMed Abstract: For the whole GFP family, a few cases, when a single mutation in the chromophore environment strongly inhibits maturation, were described. Here we study EYFP-F165G - a variant of the enhanced yellow fluorescent protein - obtained by a single F165G replacement, and demonstrated multiple fluorescent states represented by the minor emission peaks in blue and yellow ranges (~470 and ~530 nm), and the major peak at ~330 nm. The latter has been assigned to tryptophan fluorescence, quenched due to excitation energy transfer to the mature chromophore in the parental EYFP protein. EYFP-F165G crystal structure revealed two general independent routes of post-translational chemistry, resulting in two main states of the polypeptide chain with the intact chromophore forming triad (~85%) and mature chromophore (~15%). Our experiments thus highlighted important stereochemical role of the 165th position strongly affecting spectral characteristics of the protein. On the basis of the determined EYFP-F165G three-dimensional structure, new variants with ~ 2-fold improved brightness were engineered.

PubMed: 34136094
DOI: 10.1016/j.csbj.2021.05.017

Experimental method

X-RAY DIFFRACTION (2.2 Å)