PDBsum entry 4zfs

Fluorescent protein

PDB id: 4zfs

Contents

Protein chains

234 a.a.

Waters ×862

PDB id:

4zfs

Name:

Fluorescent protein

Title:

Phototoxic fluorescent protein killerorange

Structure:

Killerorange. Chain: a, b, c, d, e. Engineered: yes. Other_details: killerorange

Source:

Hydrozoa. Hydrozoans. Organism_taxid: 6074. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.01Å R-factor: 0.192 R-free: 0.236

Authors:

N.V.Pletneva,V.Z.Pletnev,S.Pletnev

Key ref:

N.V.Pletneva et al. (2015). Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore. Plos One, 10, e0145740. PubMed id: 26699366 DOI: 10.1371/journal.pone.0145740

Date:

21-Apr-15 Release date: 23-Dec-15

Protein chains

Q2TCH5  (Q2TCH5_9CNID) -  KillerRed from Anthomedusae sp. DC-2005

Seq: 237 a.a.

Struc: 234 a.a.*

* PDB and UniProt seqs differ at 8 residue positions (black crosses)

DOI no: 10.1371/journal.pone.0145740

Plos One 10:e0145740 (2015)

PubMed id: 26699366

Crystal Structure of Phototoxic Orange Fluorescent Proteins with a Tryptophan-Based Chromophore.

N.V.Pletneva, V.Z.Pletnev, K.S.Sarkisyan, D.A.Gorbachev, E.S.Egorov, A.S.Mishin, K.A.Lukyanov, Z.Dauter, S.Pletnev.

ABSTRACT

Phototoxic fluorescent proteins represent a sparse group of genetically encoded photosensitizers that could be used for precise light-induced inactivation of target proteins, DNA damage, and cell killing. Only two such GFP-based fluorescent proteins (FPs), KillerRed and its monomeric variant SuperNova, were described up to date. Here, we present a crystallographic study of their two orange successors, dimeric KillerOrange and monomeric mKillerOrange, at 1.81 and 1.57 Å resolution, respectively. They are the first orange-emitting protein photosensitizers with a tryptophan-based chromophore (Gln65-Trp66-Gly67). Same as their red progenitors, both orange photosensitizers have a water-filled channel connecting the chromophore to the β-barrel exterior and enabling transport of ROS. In both proteins, Trp66 of the chromophore adopts an unusual trans-cis conformation stabilized by H-bond with the nearby Gln159. This trans-cis conformation along with the water channel was shown to be a key structural feature providing bright orange emission and phototoxicity of both examined orange photosensitizers.