PDBsum entry 2g2s

Luminescent protein

PDB id: 2g2s

Contents

Protein chains

64 a.a. *

165 a.a. *

Metals

_MG

Waters ×325

* Residue conservation analysis

PDB id:

2g2s

Name:

Luminescent protein

Title:

Structure of s65g y66s gfp variant after spontaneous peptide hydrolysis

Structure:

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

Source:

Aequorea victoria. Organism_taxid: 6100. Gene: gfp. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

Biol. unit:

Dimer (from PQS)

Resolution:

1.20Å R-factor: 0.123 R-free: 0.174

Authors:

D.P.Barondeau

Key ref:

D.P.Barondeau et al. (2006). Understanding GFP posttranslational chemistry: structures of designed variants that achieve backbone fragmentation, hydrolysis, and decarboxylation. J Am Chem Soc, 128, 4685-4693. PubMed id: 16594705 DOI: 10.1021/ja056635l

Date:

16-Feb-06 Release date: 18-Apr-06

Protein chain

P42212  (GFP_AEQVI) -  Green fluorescent protein from Aequorea victoria

Seq: 238 a.a.

Struc: 64 a.a.*

Protein chain

P42212  (GFP_AEQVI) -  Green fluorescent protein from Aequorea victoria

Seq: 238 a.a.

Struc: 165 a.a.*

* PDB and UniProt seqs differ at 6 residue positions (black crosses)

DOI no: 10.1021/ja056635l

J Am Chem Soc 128:4685-4693 (2006)

PubMed id: 16594705

Understanding GFP posttranslational chemistry: structures of designed variants that achieve backbone fragmentation, hydrolysis, and decarboxylation.

D.P.Barondeau, C.J.Kassmann, J.A.Tainer, E.D.Getzoff.

ABSTRACT

The green fluorescent protein (GFP) creates a fluorophore out of three sequential amino acids by promoting spontaneous posttranslational modifications. Here, we use high-resolution crystallography to characterize GFP variants that not only undergo peptide backbone cyclization but additional denaturation-induced peptide backbone fragmentation, native peptide hydrolysis, and decarboxylation reactions. Our analyses indicate that architectural features that favor GFP peptide cyclization also drive peptide hydrolysis. These results are relevant for the maturation pathways of GFP homologues, such as the kindling fluorescent protein and the Kaede protein, which use backbone cleavage to red-shift the spectral properties of their chromophores. We further propose a photochemical mechanism for the decarboxylation reaction, supporting a role for the GFP protein environment in facilitating radical formation and one-electron chemistry, which may be important in activating oxygen for the oxidation step of chromophore biosynthesis. Together, our results characterize GFP posttranslational modification chemistry with implications for the energetic landscape of backbone cyclization and subsequent reactions, and for the rational design of predetermined spontaneous backbone cyclization and cleavage reactions.