PDBsum entry 2o2b

Luminescent protein

PDB id

2o2b

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Contents

			Protein chain

					234 a.a. *

			Metals

					IOD ×9

			Waters ×148

* Residue conservation analysis

PDB id:

2o2b

Links

Name:

Luminescent protein

Title:

Spectroscopic and structural study of the heterotropic linkage between halide and proton ion binding to gfp proteins: e2(gfp)-i complex

Structure:

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

Source:

Aequorea victoria. Organism_taxid: 6100. Gene: gfp. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.94Å

R-factor:

0.184

R-free:

0.234

Authors:

G.Garau

Key ref:

D.Arosio et al. (2007). Spectroscopic and structural study of proton and halide ion cooperative binding to gfp. Biophys J, 93, 232-244. PubMed id: 17434942

Date:

29-Nov-06

Release date:

15-May-07

PROCHECK

	Headers

	References

Protein chain

P42212 (GFP_AEQVI) - Green fluorescent protein from Aequorea victoria

Seq: Struc:					238 a.a. 234 a.a.*

Key:

PfamA domain

Secondary structure

CATH domain

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

	Biophys J 93:232-244 (2007)
PubMed id: 17434942

Spectroscopic and structural study of proton and halide ion cooperative binding to gfp.
D.Arosio, G.Garau, F.Ricci, L.Marchetti, R.Bizzarri, R.Nifosì, F.Beltram.

ABSTRACT

This study reports the influence of halogens on fluorescence properties of the Aequorea victoria Green Fluorescent Protein variant S65T/T203Y (E(2)GFP). Halide binding forms a specific nonfluorescent complex generating a substantial drop of the fluorescence via static quenching. Spectroscopic analysis under different solution conditions reveals high halogen affinity, which is strongly dependent on the pH. This evidences the presence in E(2)GFP of interacting binding sites for halide ions and for protons. Thermodynamic link and cooperative interaction are assessed demonstrating that binding of one halide ion is associated with the binding of one proton in a cooperative fashion with the formation, in the pH range 4.5-10, of a single fully protonated E(2)GFP.halogen complex. To resolve the structural determinants of E(2)GFP sensitivity to halogens, high-resolution crystallographic structures were obtained for the halide-free and I(-), Br(-), and Cl(-) bound E(2)GFP. Remarkably the first high-resolution (1.4 A) crystallographic structure of a chloride-bound GFP is reported. The chloride ion occupies a specific and unique binding pocket in direct contact (3.4 A) with the chromophore imidazolidinone aromatic ring. Unanticipated flexibility, strongly modulated by halide ion interactions, is observed in the region surrounding the chromophore. Furthermore molecular dynamics simulations identified E222 residue (along with the chromophore Y66 residue) being in the protonated state when E(2)GFP.halogen complex is formed. The impact of these results on high-sensitivity biosensor design will be discussed.

Literature references that cite this PDB file's key reference

PubMed id

Reference

21347442

T.P.Burghardt (2011).
Single molecule fluorescence image patterns linked to dipole orientation and axial position: application to Myosin cross-bridges in muscle fibers.

PLoS One, 6, e16772.

20581829

D.Arosio, F.Ricci, L.Marchetti, R.Gualdani, L.Albertazzi, and F.Beltram (2010).
Simultaneous intracellular chloride and pH measurements using a GFP-based sensor.

Nat Methods, 7, 516-518.

20693533

L.Marchetti, L.Comelli, B.D'Innocenzo, L.Puzzi, S.Luin, D.Arosio, M.Calvello, R.Mendoza-Maldonado, F.Peverali, F.Trovato, S.Riva, G.Biamonti, G.Abdurashidova, F.Beltram, and A.Falaschi (2010).
Homeotic proteins participate in the function of human-DNA replication origins.

Nucleic Acids Res, 38, 8105-8119.

19901033

S.T.Hsu, G.Blaser, C.Behrens, L.D.Cabrita, C.M.Dobson, and S.E.Jackson (2010).
Folding study of Venus reveals a strong ion dependence of its yellow fluorescence under mildly acidic conditions.

J Biol Chem, 285, 4859-4869.

19771331

H.E.Seward, and C.R.Bagshaw (2009).
The photochemistry of fluorescent proteins: implications for their biological applications.

Chem Soc Rev, 38, 2842-2851.

19034433

R.Bizzarri, M.Serresi, S.Luin, and F.Beltram (2009).
Green fluorescent protein based pH indicators for in vivo use: a review.

Anal Bioanal Chem, 393, 1107-1122.

19771338

S.T.Hsu, G.Blaser, and S.E.Jackson (2009).
The folding, stability and conformational dynamics of beta-barrel fluorescent proteins.

Chem Soc Rev, 38, 2951-2965.

19127992

T.P.Burghardt, J.Li, and K.Ajtai (2009).
Single myosin lever arm orientation in a muscle fiber detected with photoactivatable GFP.

Biochemistry, 48, 754-765.

18818813

S.Girardo, M.Cecchini, F.Beltram, R.Cingolani, and D.Pisignano (2008).
Polydimethylsiloxane-LiNbO3 surface acoustic wave micropump devices for fluid control into microchannels.

Lab Chip, 8, 1557-1563.

18071586

N.Tansila, T.Tantimongcolwat, C.Isarankura-Na-Ayudhya, C.Nantasenamat, and V.Prachayasittikul (2007).
Rational design of analyte channels of the green fluorescent protein for biosensor applications.

Int J Biol Sci, 3, 463-470.

The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.