Reviewed, UniProtKB/Swiss-Prot P42212 (GFP_AEQVI)

Names and origin

Protein names

Recommended name:
    Green fluorescent protein

Gene names

Name:

GFP

Organism

Aequorea victoria (Jellyfish)

Taxonomic identifier

6100 [NCBI]

Taxonomic lineage

Eukaryota › Metazoa › Cnidaria › Hydrozoa › Hydroida › Leptomedusae › Aequoreidae › Aequorea

Protein attributes

Sequence length	238 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin.
Subunit structure	Monomer.
Tissue specificity	Photocytes.
Post-translational modification	Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.
Biotechnological use	Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.
Sequence similarities	Belongs to the GFP family.
Biophysicochemical properties	Absorption: Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm. Abs(max)=395 nm

Ontologies

Keywords
Biological process	Luminescence
Ligand	Chromophore
Molecular function	Photoprotein
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological process	bioluminescence Ref.2 Traceable author statement. Source: UniProtKB generation of precursor metabolites and energy Ref.2 Traceable author statement. Source: UniProtKB
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Molecule processing

Chain

1 – 238

238

Green fluorescent protein

Amino acid modifications

Modified residue

66

1

(Z)-2,3-didehydrotyrosine

Cross-link

65 ↔ 67

5-imidazolinone (Ser-Gly)

Natural variations

Natural variant

100

1

F → Y

Natural variant

108

1

T → S

Natural variant

141

1

L → M

Natural variant

219

1

V → I

Experimental info

Mutagenesis

65

1

S → G in mut3; highly fluorescent mutant when excited at 488 nm; when associated with A-72

Mutagenesis

65

1

S → T: Increases fluoresence, photostability and shift the major exitation peak to 488 nm

Mutagenesis

72

1

S → A in mut3; highly fluorescent mutant when excited at 488 nm; when associated with G-65

Sequence conflict

2

1

S → G in CAA65278. Ref.3

Sequence conflict

25

1

H → Q in CAA65278. Ref.2

Sequence conflict

80

1

Q → R in CAA65278. Ref.3

Sequence conflict

157

1

Q → P in AAA58246. Ref.2

Sequence conflict

172

1

E → K in AAA58246. Ref.2

Secondary structure

1

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238

Helix Strand Turn

Details...

Helix

5 – 8

4

Beta strand

12 – 22

11

Beta strand

25 – 36

12

Helix

37 – 39

3

Beta strand

41 – 48

8

Helix

57 – 60

4

Turn

61 – 63

3

Helix

69 – 71

3

Helix

76 – 81

6

Helix

83 – 86

4

Turn

87 – 90

4

Beta strand

92 – 100

9

Beta strand

105 – 115

11

Beta strand

118 – 129

12

Turn

135 – 139

5

Beta strand

149 – 155

7

Helix

156 – 158

3

Beta strand

160 – 171

12

Beta strand

176 – 191

16

Beta strand

198 – 208

11

Beta strand

215 – 228

14

Sequences

	Sequence		Length	Mass (Da)
	P42212-1 [UniParc]. Last modified November 1, 1995. Version 1. Checksum: EA5A6F21FBFB6E05	FASTA	238	26,886
	10 20 30 40 50 60 MSKGEELFTG VVPILVELDG DVNGHKFSVS GEGEGDATYG KLTLKFICTT GKLPVPWPTL 70 80 90 100 110 120 VTTFSYGVQC FSRYPDHMKQ HDFFKSAMPE GYVQERTIFF KDDGNYKTRA EVKFEGDTLV 130 140 150 160 170 180 NRIELKGIDF KEDGNILGHK LEYNYNSHNV YIMADKQKNG IKVNFKIRHN IEDGSVQLAD 190 200 210 220 230 HYQQNTPIGD GPVLLPDNHY LSTQSALSKD PNEKRDHMVL LEFVTAAGIT HGMDELYK

References

[1]	"Primary structure of the Aequorea victoria green-fluorescent protein." Prasher D.C., Eckenrode V.K., Ward W.W., Prendergast F.G., Cormier M.J. Gene 111:229-233(1992) [PubMed: 1347277] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], PARTIAL PROTEIN SEQUENCE, VARIANTS TYR-100; SER-108; MET-141 AND ILE-219.
[2]	"Aequorea green fluorescent protein. Expression of the gene and fluorescence characteristics of the recombinant protein." Inouye S., Tsuji F.I. FEBS Lett. 341:277-280(1994) [PubMed: 8137953] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA].
[3]	"Enhanced expression in tobacco of the gene encoding green fluorescent protein by modification of its codon usage." Rouwendal G.J.A., Mendes O., Wolbert E.J.H., de Boer A.D. Plant Mol. Biol. 33:989-999(1997) [PubMed: 9154981] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA].
[4]	"FACS-optimized mutants of the green fluorescent protein (GFP)." Cormack B.P., Valdivia R.H., Falkow S. Gene 173:33-38(1996) [PubMed: 8707053] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], MUTAGENESIS OF SER-65 AND SER-72.
[5]	"Chemical structure of the hexapeptide chromophore of the Aequorea green-fluorescent protein." Cody C.W., Prasher D.C., Westler W.M., Prendergast F.G., Ward W.W. Biochemistry 32:1212-1218(1993) [PubMed: 8448132] [Abstract] Cited for: CHROMOPHORE.
[6]	"A molecular thermometer based on fluorescent protein blinking." Wong F.H., Banks D.S., Abu-Arish A., Fradin C. J. Am. Chem. Soc. 129:10302-10303(2007) [PubMed: 17685514] [Abstract] Cited for: BIOTECHNOLOGY.
[7]	"Crystal structure of the Aequorea victoria green fluorescent protein." Ormoe M., Cubitt A.B., Kallio K., Gross L.A., Tsien R.Y., Remington S.J. Science 273:1392-1395(1996) [PubMed: 8703075] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF MUTANT THR-65.
[8]	"The molecular structure of green fluorescent protein." Yang F., Moss L.G., Phillips G.N. Jr. Nat. Biotechnol. 14:1246-1251(1996) [PubMed: 9631087] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS).
[9]	"Structural basis of spectral shifts in the yellow-emission variants of green fluorescent protein." Wachter R.M., Elsliger M.-A., Kallio K., Hanson G.T., Remington S.J. Structure 6:1267-1277(1998) [PubMed: 9782051] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) OF MUTANT WITH YELLOW EMISSION.
[10]	"Structural and spectral response of green fluorescent protein variants to changes in pH." Elsliger M.-A., Wachter R.M., Hanson G.T., Kallio K., Remington S.J. Biochemistry 38:5296-5301(1999) [PubMed: 10220315] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
+	Additional computationally mapped references.

Web resources

Protein Spotlight

The greenest of us all - Issue 11 of June 2001

Wikipedia

Green fluorescent protein entry

Cross-references

Sequence databases

EMBLGenBankDDBJ

M62654 mRNA. Translation: AAA27722.1.
M62653 mRNA. Translation: AAA27721.1.
L29345 mRNA. Translation: AAA58246.1.
X96418 mRNA. Translation: CAA65278.1.
U73901 Genomic DNA. Translation: AAB18957.1.

PIR

JQ1514. JS0692.

3D structure databases

PDBMSD

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1B9C	X-ray	2.40	A/B/C/D	2-238	[»]
1BFP	X-ray	2.10	A	1-238	[»]
1C4F	X-ray	2.25	A	1-238	[»]
1CV7	X-ray	2.50	A	1-228	[»]
1EMA	X-ray	1.90	A	1-238	[»]
1EMB	X-ray	2.13	A	1-238	[»]
1EMC	X-ray	2.30	A/B/C/D	2-237	[»]
1EME	X-ray	2.50	A	2-237	[»]
1EMF	X-ray	2.40	A	2-238	[»]
1EMG	X-ray	2.00	A	1-238	[»]
1EMK	X-ray	2.10	A	2-237	[»]
1EML	X-ray	2.30	A	2-237	[»]
1EMM	X-ray	2.30	A	2-238	[»]
1F09	X-ray	2.14	A	1-238	[»]
1F0B	X-ray	2.10	A	1-238	[»]
1GFL	X-ray	1.90	A/B	2-238	[»]
1H6R	X-ray	1.50	A/B/C	1-238	[»]
1HCJ	X-ray	1.80	A/B/C/D	1-238	[»]
1HUY	X-ray	2.20	A	2-238	[»]
1JBY	X-ray	1.80	A	1-238	[»]
1JBZ	X-ray	1.50	A	1-238	[»]
1JC0	X-ray	2.00	A/B/C	1-238	[»]
1JC1	X-ray	1.90	A/B/C	1-238	[»]
1KP5	X-ray	2.60	A/B	3-238	[»]
1KYP	X-ray	1.35	A	2-238	[»]
1KYR	X-ray	1.50	A	2-238	[»]
1KYS	X-ray	1.44	A	2-238	[»]
1MYW	X-ray	2.20	A	1-238	[»]
1Q4A	X-ray	1.45	A	1-238	[»]
1Q4B	X-ray	1.48	A	1-238	[»]
1Q4C	X-ray	1.55	A	1-238	[»]
1Q4D	X-ray	1.58	A	1-238	[»]
1Q4E	X-ray	1.38	A	1-238	[»]
1Q73	X-ray	1.60	A	1-238	[»]
1QXT	X-ray	2.00	A	2-229	[»]
1QY3	X-ray	2.00	A	1-229	[»]
1QYF	X-ray	1.50	A	2-229	[»]
1QYO	X-ray	1.80	A	2-238	[»]
1QYQ	X-ray	1.80	A	2-238	[»]
1RM9	X-ray	2.90	A	2-238	[»]
1RMM	X-ray	1.90	A	2-229	[»]
1RMO	X-ray	1.80	A	2-238	[»]
1RMP	X-ray	3.00	A	2-229	[»]
1RRX	X-ray	2.10	A	2-229	[»]
1W7S	X-ray	1.85	A/B/C/D	1-238	[»]
1W7T	X-ray	1.85	A/B/C/D	1-238	[»]
1W7U	X-ray	1.85	A/B/C/D	1-238	[»]
1YFP	X-ray	2.50	A/B	3-229	[»]
1YHG	X-ray	2.50	A/B	2-238	[»]
1YHH	X-ray	1.50	A	2-238	[»]
1YHI	X-ray	1.90	A	2-238	[»]
1YJ2	X-ray	1.50	A	2-238	[»]
1YJF	X-ray	1.35	A	2-238	[»]
1Z1P	X-ray	2.00	A	1-238	[»]
1Z1Q	X-ray	1.50	A	1-238	[»]
2AH8	X-ray	2.24	A/B	1-238	[»]
2AHA	X-ray	1.98	A/B	1-238	[»]
2AWJ	X-ray	1.60	A	2-229	[»]
2AWK	X-ray	1.15	A	2-229	[»]
2AWL	X-ray	1.85	A	2-229	[»]
2AWM	X-ray	1.70	A	2-229	[»]
2B3P	X-ray	1.40	A	1-238	[»]
2B3Q	X-ray	2.30	A/B/C/D	1-238	[»]
2DUE	X-ray	1.24	A	1-238	[»]
2DUF	X-ray	1.50	A	1-238	[»]
2DUG	X-ray	1.40	A	1-238	[»]
2DUH	X-ray	1.20	A	1-238	[»]
2DUI	X-ray	1.36	A	1-238	[»]
2EMD	X-ray	2.00	A	2-238	[»]
2EMN	X-ray	2.30	A	2-238	[»]
2EMO	X-ray	2.60	A	2-238	[»]
2FWQ	X-ray	1.40	A	2-238	[»]
2FZU	X-ray	1.25	A	2-238	[»]
2G16	X-ray	2.00	A	2-64	[»]
			B	68-238	[»]
2G2S	X-ray	1.20	A	2-65	[»]
			B	67-238	[»]
2G3D	X-ray	1.35	A	2-65	[»]
			B	66-238	[»]