Tags

Type your tag names separated by a space and hit enter

Rational Engineering of Photoconvertible Fluorescent Proteins for Dual-Color Fluorescence Nanoscopy Enabled by a Triplet-State Mechanism of Primed Conversion.
Angew Chem Int Ed Engl 2017; 56(38):11628-11633AC

Abstract

Green-to-red photoconvertible fluorescent proteins (pcFPs) are powerful tools for super-resolution localization microscopy and protein tagging. Recently, they have been found to undergo efficient photoconversion not only by the traditional 400-nm illumination but also by an alternative method termed primed conversion, employing dual wavelength illumination with blue and far-red/near-infrared light. Primed conversion has been reported only for Dendra2 and its mechanism has remained elusive. Here, we uncover the molecular mechanism of primed conversion by reporting the intermediate "primed" state to be a triplet dark state formed by intersystem crossing. We show that formation of this state can be influenced by the introduction of serine or threonine at sequence position 69 (Eos notation) and use this knowledge to create "pr"- (for primed convertible) variants of most known green-to-red pcFPs.

Authors+Show Affiliations

Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058, Basel, Switzerland. Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA, 20147, USA.Institute of Applied Physics, Karlsruhe Institute of Technology (KIT), 76131, Karlsruhe, Germany.Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058, Basel, Switzerland.Institute of Applied Physics, Karlsruhe Institute of Technology (KIT), 76131, Karlsruhe, Germany.Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA, 20147, USA.Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA, 20147, USA.Institute of Applied Physics, Karlsruhe Institute of Technology (KIT), 76131, Karlsruhe, Germany. Institute of Toxicology and Genetics, Karlsruhe Institute of Technology (KIT), 76344, Eggenstein-Leopoldshafen, Germany. Institute of Nanotechnology, Karlsruhe Institute of Technology (KIT), 76344, Eggenstein-Leopoldshafen, Germany. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058, Basel, Switzerland.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28661566

Citation

Mohr, Manuel Alexander, et al. "Rational Engineering of Photoconvertible Fluorescent Proteins for Dual-Color Fluorescence Nanoscopy Enabled By a Triplet-State Mechanism of Primed Conversion." Angewandte Chemie (International Ed. in English), vol. 56, no. 38, 2017, pp. 11628-11633.
Mohr MA, Kobitski AY, Sabater LR, et al. Rational Engineering of Photoconvertible Fluorescent Proteins for Dual-Color Fluorescence Nanoscopy Enabled by a Triplet-State Mechanism of Primed Conversion. Angew Chem Int Ed Engl. 2017;56(38):11628-11633.
Mohr, M. A., Kobitski, A. Y., Sabater, L. R., Nienhaus, K., Obara, C. J., Lippincott-Schwartz, J., ... Pantazis, P. (2017). Rational Engineering of Photoconvertible Fluorescent Proteins for Dual-Color Fluorescence Nanoscopy Enabled by a Triplet-State Mechanism of Primed Conversion. Angewandte Chemie (International Ed. in English), 56(38), pp. 11628-11633. doi:10.1002/anie.201706121.
Mohr MA, et al. Rational Engineering of Photoconvertible Fluorescent Proteins for Dual-Color Fluorescence Nanoscopy Enabled By a Triplet-State Mechanism of Primed Conversion. Angew Chem Int Ed Engl. 2017 09 11;56(38):11628-11633. PubMed PMID: 28661566.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rational Engineering of Photoconvertible Fluorescent Proteins for Dual-Color Fluorescence Nanoscopy Enabled by a Triplet-State Mechanism of Primed Conversion. AU - Mohr,Manuel Alexander, AU - Kobitski,Andrei Yu, AU - Sabater,Lluc Rullan, AU - Nienhaus,Karin, AU - Obara,Christopher John, AU - Lippincott-Schwartz,Jennifer, AU - Nienhaus,Gerd Ulrich, AU - Pantazis,Periklis, Y1 - 2017/07/11/ PY - 2017/06/15/received PY - 2017/7/1/pubmed PY - 2019/3/22/medline PY - 2017/6/30/entrez KW - biophysics KW - fluorescent proteins KW - primed conversion KW - super-resolution microscopy KW - triplet states SP - 11628 EP - 11633 JF - Angewandte Chemie (International ed. in English) JO - Angew. Chem. Int. Ed. Engl. VL - 56 IS - 38 N2 - Green-to-red photoconvertible fluorescent proteins (pcFPs) are powerful tools for super-resolution localization microscopy and protein tagging. Recently, they have been found to undergo efficient photoconversion not only by the traditional 400-nm illumination but also by an alternative method termed primed conversion, employing dual wavelength illumination with blue and far-red/near-infrared light. Primed conversion has been reported only for Dendra2 and its mechanism has remained elusive. Here, we uncover the molecular mechanism of primed conversion by reporting the intermediate "primed" state to be a triplet dark state formed by intersystem crossing. We show that formation of this state can be influenced by the introduction of serine or threonine at sequence position 69 (Eos notation) and use this knowledge to create "pr"- (for primed convertible) variants of most known green-to-red pcFPs. SN - 1521-3773 UR - https://www.unboundmedicine.com/medline/citation/28661566/Rational_Engineering_of_Photoconvertible_Fluorescent_Proteins_for_Dual_Color_Fluorescence_Nanoscopy_Enabled_by_a_Triplet_State_Mechanism_of_Primed_Conversion_ L2 - https://doi.org/10.1002/anie.201706121 DB - PRIME DP - Unbound Medicine ER -