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Structural basis for bathochromic shift of fluorescence in far-red fluorescent proteins eqFP650 and eqFP670.
Acta Crystallogr D Biol Crystallogr 2012; 68(Pt 9):1088-97AC

Abstract

The crystal structures of the far-red fluorescent proteins (FPs) eqFP650 (λ(ex)(max)/λ(em)(max) 592/650 nm) and eqFP670 (λ(ex)(max)/λ(em)(max) 605/670 nm), the successors of the far-red FP Katushka (λ(ex)(max)/λ(em)(max) 588/635 nm), have been determined at 1.8 and 1.6 Å resolution, respectively. An examination of the structures demonstrated that there are two groups of changes responsible for the bathochromic shift of excitation/emission bands of these proteins relative to their predecessor. The first group of changes resulted in an increase of hydrophilicity at the acylimine site of the chromophore due to the presence of one and three water molecules in eqFP650 and eqFP670, respectively. These water molecules provide connection of the chromophore with the protein scaffold via hydrogen bonds causing an ~15 nm bathochromic shift of the eqFP650 and eqFP670 emission bands. The second group of changes observed in eqFP670 arises from substitution of both Ser143 and Ser158 by asparagines. Asn143 and Asn158 of eqFP670 are hydrogen bonded with each other, as well as with the protein scaffold and with the p-hydroxyphenyl group of the chromophore, resulting in an additional ~20 nm bathochromic shift of the eqFP670 emission band as compared to eqFP650. The role of the observed structural changes was verified by mutagenesis.

Authors+Show Affiliations

Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, National Cancer Institute, Argonne, IL 60439, USA. pletnevs@mail.nih.govNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

22948909

Citation

Pletnev, Sergei, et al. "Structural Basis for Bathochromic Shift of Fluorescence in Far-red Fluorescent Proteins eqFP650 and EqFP670." Acta Crystallographica. Section D, Biological Crystallography, vol. 68, no. Pt 9, 2012, pp. 1088-97.
Pletnev S, Pletneva NV, Souslova EA, et al. Structural basis for bathochromic shift of fluorescence in far-red fluorescent proteins eqFP650 and eqFP670. Acta Crystallogr D Biol Crystallogr. 2012;68(Pt 9):1088-97.
Pletnev, S., Pletneva, N. V., Souslova, E. A., Chudakov, D. M., Lukyanov, S., Wlodawer, A., ... Pletnev, V. (2012). Structural basis for bathochromic shift of fluorescence in far-red fluorescent proteins eqFP650 and eqFP670. Acta Crystallographica. Section D, Biological Crystallography, 68(Pt 9), pp. 1088-97. doi:10.1107/S0907444912020598.
Pletnev S, et al. Structural Basis for Bathochromic Shift of Fluorescence in Far-red Fluorescent Proteins eqFP650 and EqFP670. Acta Crystallogr D Biol Crystallogr. 2012;68(Pt 9):1088-97. PubMed PMID: 22948909.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structural basis for bathochromic shift of fluorescence in far-red fluorescent proteins eqFP650 and eqFP670. AU - Pletnev,Sergei, AU - Pletneva,Nadya V, AU - Souslova,Ekaterina A, AU - Chudakov,Dmitry M, AU - Lukyanov,Sergey, AU - Wlodawer,Alexander, AU - Dauter,Zbigniew, AU - Pletnev,Vladimir, Y1 - 2012/08/18/ PY - 2012/03/30/received PY - 2012/05/07/accepted PY - 2012/9/6/entrez PY - 2012/9/6/pubmed PY - 2013/5/2/medline SP - 1088 EP - 97 JF - Acta crystallographica. Section D, Biological crystallography JO - Acta Crystallogr. D Biol. Crystallogr. VL - 68 IS - Pt 9 N2 - The crystal structures of the far-red fluorescent proteins (FPs) eqFP650 (λ(ex)(max)/λ(em)(max) 592/650 nm) and eqFP670 (λ(ex)(max)/λ(em)(max) 605/670 nm), the successors of the far-red FP Katushka (λ(ex)(max)/λ(em)(max) 588/635 nm), have been determined at 1.8 and 1.6 Å resolution, respectively. An examination of the structures demonstrated that there are two groups of changes responsible for the bathochromic shift of excitation/emission bands of these proteins relative to their predecessor. The first group of changes resulted in an increase of hydrophilicity at the acylimine site of the chromophore due to the presence of one and three water molecules in eqFP650 and eqFP670, respectively. These water molecules provide connection of the chromophore with the protein scaffold via hydrogen bonds causing an ~15 nm bathochromic shift of the eqFP650 and eqFP670 emission bands. The second group of changes observed in eqFP670 arises from substitution of both Ser143 and Ser158 by asparagines. Asn143 and Asn158 of eqFP670 are hydrogen bonded with each other, as well as with the protein scaffold and with the p-hydroxyphenyl group of the chromophore, resulting in an additional ~20 nm bathochromic shift of the eqFP670 emission band as compared to eqFP650. The role of the observed structural changes was verified by mutagenesis. SN - 1399-0047 UR - https://www.unboundmedicine.com/medline/citation/22948909/Structural_basis_for_bathochromic_shift_of_fluorescence_in_far_red_fluorescent_proteins_eqFP650_and_eqFP670_ L2 - http://scripts.iucr.org/cgi-bin/paper?S0907444912020598 DB - PRIME DP - Unbound Medicine ER -