PDBsum entry 3lvc

Fluorescent protein

PDB id: 3lvc

Contents

Protein chains

227 a.a. *

Ligands

GOL ×5

Waters ×486

* Residue conservation analysis

PDB id:

3lvc

Name:

Fluorescent protein

Title:

Crystal structure of gfp-like protein acegfp_g222e (a. Coerulescens). Colorless form.

Structure:

Green fluorescent protein. Chain: a, b. Engineered: yes. Mutation: yes

Source:

Aequorea coerulescens. Organism_taxid: 210840. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.14Å R-factor: 0.139 R-free: 0.156

Authors:

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

Key ref:

N.V.Pletneva et al. (2010). Structural evidence for a dehydrated intermediate in green fluorescent protein chromophore biosynthesis. J Biol Chem, 285, 15978-15984. PubMed id: 20220148

Date:

19-Feb-10 Release date: 09-Mar-10

Protein chains

Q6YGZ0  (Q6YGZ0_9CNID) -  Green fluorescent protein from Aequorea coerulescens

Seq: 238 a.a.

Struc: 227 a.a.*

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

J Biol Chem 285:15978-15984 (2010)

PubMed id: 20220148

Structural evidence for a dehydrated intermediate in green fluorescent protein chromophore biosynthesis.

N.V.Pletneva, V.Z.Pletnev, K.A.Lukyanov, N.G.Gurskaya, E.A.Goryacheva, V.I.Martynov, A.Wlodawer, Z.Dauter, S.Pletnev.

ABSTRACT

The acGFPL is the first-identified member of a novel, colorless and non-fluorescent group of green fluorescent protein (GFP)-like proteins. Its mutant aceGFP, with Gly replacing the invariant catalytic Glu-222, demonstrates a relatively fast maturation rate and bright green fluorescence (lambda(ex) = 480 nm, lambda(em) = 505 nm). The reverse G222E single mutation in aceGFP results in the immature, colorless variant aceGFP-G222E, which undergoes irreversible photoconversion to a green fluorescent state under UV light exposure. Here we present a high resolution crystallographic study of aceGFP and aceGFP-G222E in the immature and UV-photoconverted states. A unique and striking feature of the colorless aceGFP-G222E structure is the chromophore in the trapped intermediate state, where cyclization of the protein backbone has occurred, but Tyr-66 still stays in the native, non-oxidized form, with C(alpha) and C(beta) atoms in the sp(3) hybridization. This experimentally observed immature aceGFP-G222E structure, characterized by the non-coplanar arrangement of the imidazolone and phenolic rings, has been attributed to one of the intermediate states in the GFP chromophore biosynthesis. The UV irradiation (lambda = 250-300 nm) of aceGFP-G222E drives the chromophore maturation further to a green fluorescent state, characterized by the conventional coplanar bicyclic structure with the oxidized double Tyr-66 C(alpha)=C(beta) bond and the conjugated system of pi-electrons. Structure-based site-directed mutagenesis has revealed a critical role of the proximal Tyr-220 in the observed effects. In particular, an alternative reaction pathway via Tyr-220 rather than conventional wild type Glu-222 has been proposed for aceGFP maturation.