PDBsum entry 4hq9

Fluorescent protein

PDB id: 4hq9

Contents

Protein chains

(+ 0 more) 217 a.a.

Ligands

PEG ×4

PGE

PG4 ×2

Waters ×601

PDB id:

4hq9

Name:

Fluorescent protein

Title:

Crystal structure of a green-to-red photoconvertible dronpa, pcdronpa in the green-off-state

Structure:

Fluorescent protein dronpa. Chain: a, b, c, d, e, f. Synonym: green fluorescent protein. Engineered: yes. Mutation: yes

Source:

Echinophyllia. Organism_taxid: 301887. Strain: sc22. Gene: dronpa. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.07Å R-factor: 0.191 R-free: 0.235

Authors:

N.Nguyen Bich,L.Van Meervelt

Key ref:

B.Moeyaert et al. (2014). Green-to-red photoconvertible Dronpa mutant for multimodal super-resolution fluorescence microscopy. Acs Nano, 8, 1664-1673. PubMed id: 24410188 DOI: 10.1021/nn4060144

Date:

25-Oct-12 Release date: 13-Nov-13

Protein chains

Q5TLG6  (Q5TLG6_9CNID) -  Fluorescent protein Dronpa from Echinophyllia sp. SC22

Seq: 224 a.a.

Struc: 217 a.a.*

* PDB and UniProt seqs differ at 5 residue positions (black crosses)

DOI no: 10.1021/nn4060144

Acs Nano 8:1664-1673 (2014)

PubMed id: 24410188

Green-to-red photoconvertible Dronpa mutant for multimodal super-resolution fluorescence microscopy.

B.Moeyaert, N.Nguyen Bich, E.De Zitter, S.Rocha, K.Clays, H.Mizuno, L.van Meervelt, J.Hofkens, P.Dedecker.

ABSTRACT

Advanced imaging techniques crucially depend on the labels used. In this work, we present the structure-guided design of a fluorescent protein that displays both reversibly photochromic and green-to-red photoconversion behavior. We first designed ffDronpa, a mutant of the photochromic fluorescent protein Dronpa that matures up to three times faster while retaining its interesting photochromic features. Using a combined evolutionary and structure-driven rational design strategy, we developed a green-to-red photoconvertible ffDronpa mutant, called pcDronpa, and explored different optimization strategies that resulted in its improved version, pcDronpa2. This fluorescent probe combines a high brightness with low photobleaching and photoblinking. We herein show that, despite its tetrameric nature, pcDronpa2 allows for multimodal subdiffraction imaging by sequentially imaging a given sample using both super-resolution fluctuation imaging and localization microscopy.