BMRBj - BMREntryMaster

Found 1 Document (1.11 seconds)

BMRB: 5666

1GFL

1H, 15N and 13C backbone assignment of the Green Fluorescent Protein (GFP)

Authors

Khan, F., Stott, K., Jackson, S.

Assembly

truncated GFPuv

Entity

1. truncated GFPuv (polymer, Thiol state: all free), 229 monomers, 25646.46 Da Detail

GSKGEELFTG VVPILVELDG DVNGHKFSVS GEGEGDATYG KLTLKFICTT GKLPVPWPTL VTTFSYGVQC FSRYPDHMKR HDFFKSAMPE GYVQERTISF KDDGNYKTRA EVKFEGDTLV NRIELKGIDF KEDGNILGHK LEYNYNSHNV YITADKQKNG IKANFKIRHN IEDGSVQLAD HYQQNTPIGD GPVLLPDNHY LSTQSALSKD PNEKRDHMVL LEFVTAAGI

Formula weight

25646.46 Da

Entity Connection

covalent 3 Detail

ID	Type	Value order	Atom ID 1	Atom ID 2
1	covalent	doub	1:SER65:C	1:TYR66:N
2	covalent	sing	1:SER65:C	1:GLY67:N
3	covalent	doub	1:TYR66:CA	1:TYR66:CB

Source organism

Aequorea victoria

Exptl. method

NMR

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 93.0 %, Completeness: 41.1 %, Completeness (bb): 74.2 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	41.1 % (1099 of 2671)	20.7 % (287 of 1387)	58.7 % (613 of 1044)	82.9 % (199 of 240)
Backbone	74.2 % (1005 of 1354)	46.0 % (216 of 470)	89.0 % (592 of 665)	90.0 % (197 of 219)
Sidechain	18.4 % (280 of 1524)	7.7 % (71 of 917)	35.3 % (207 of 586)	9.5 % (2 of 21)
Aromatic	2.4 % (6 of 248)	4.0 % (5 of 124)	0.0 % (0 of 123)	100.0 % (1 of 1)
Methyl	9.9 % (23 of 232)	6.0 % (7 of 116)	13.8 % (16 of 116)

1. Green Fluorescent Protein

Show sample condition

Sample #1

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
1	Green Fluorescent Protein	[U-95% 13C; U-95% 15N]	1.0 mM
2	sodium chloride		137 mM
3	potassium chloride		2.7 mM
4	sodium phosphate		6.0 mM

Sample #2

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
5	Green Fluorescent Protein	[U-95% 13C; U-95% 15N; U-75% 2H]	1.0 mM
6	sodium chloride		137 mM
7	potassium chloride		2.7 mM
8	sodium phosphate		6.0 mM

LACS Plot; CA

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	null	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	null	indirect	0.1013291

Referencing offset: -0.01 ppm, Outliers: 3 Detail

LACS Plot; CB

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	null	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	null	indirect	0.1013291

Referencing offset: -0.01 ppm, Outliers: 3 Detail

LACS Plot; CO

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	null	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	null	indirect	0.1013291

Referencing offset: 0.44 ppm, Outliers: 1 Detail

Release date

2003-01-22

Citation

Letter to the Editor: 1H, 15N and 13C backbone assignment of the Green Fluorescent Protein (GFP)
Khan, F., Stott, K., Jackson, S.
J. Biomol. NMR (2003), 26, 281-282

Related entities 1. truncated GFPuv, : 1 : 1 : 106 entities Detail

Interaction partners 1. truncated GFPuv, : 1 interactors Detail

More details for 1 interactors identified by 1 citations

Experiments performed 5 experiments Detail

1 HNCA

Instrument

Model Bruker DRX (500 MHz)

Sample #1

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
1	Green Fluorescent Protein	[U-95% 13C; U-95% 15N]	1.0 mM
2	sodium chloride		137 mM
3	potassium chloride		2.7 mM
4	sodium phosphate		6.0 mM

Sample #2

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
5	Green Fluorescent Protein	[U-95% 13C; U-95% 15N; U-75% 2H]	1.0 mM
6	sodium chloride		137 mM
7	potassium chloride		2.7 mM
8	sodium phosphate		6.0 mM

2 HN(CO)CA

Instrument

Model Bruker DRX (500 MHz)

Sample #1

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
1	Green Fluorescent Protein	[U-95% 13C; U-95% 15N]	1.0 mM
2	sodium chloride		137 mM
3	potassium chloride		2.7 mM
4	sodium phosphate		6.0 mM

Sample #2

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
5	Green Fluorescent Protein	[U-95% 13C; U-95% 15N; U-75% 2H]	1.0 mM
6	sodium chloride		137 mM
7	potassium chloride		2.7 mM
8	sodium phosphate		6.0 mM

3 HNCACB

Instrument

Model Bruker DRX (500 MHz)

Sample #1

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
1	Green Fluorescent Protein	[U-95% 13C; U-95% 15N]	1.0 mM
2	sodium chloride		137 mM
3	potassium chloride		2.7 mM
4	sodium phosphate		6.0 mM

Sample #2

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
5	Green Fluorescent Protein	[U-95% 13C; U-95% 15N; U-75% 2H]	1.0 mM
6	sodium chloride		137 mM
7	potassium chloride		2.7 mM
8	sodium phosphate		6.0 mM

4 HN(CO)CACB

Instrument

Model Bruker DRX (500 MHz)

Sample #1

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
1	Green Fluorescent Protein	[U-95% 13C; U-95% 15N]	1.0 mM
2	sodium chloride		137 mM
3	potassium chloride		2.7 mM
4	sodium phosphate		6.0 mM

Sample #2

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
5	Green Fluorescent Protein	[U-95% 13C; U-95% 15N; U-75% 2H]	1.0 mM
6	sodium chloride		137 mM
7	potassium chloride		2.7 mM
8	sodium phosphate		6.0 mM

5 HNCO

Instrument

Model Bruker DRX (500 MHz)

Sample #1

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
1	Green Fluorescent Protein	[U-95% 13C; U-95% 15N]	1.0 mM
2	sodium chloride		137 mM
3	potassium chloride		2.7 mM
4	sodium phosphate		6.0 mM

Sample #2

Temperature 310 (±0.5) K, pH 7.2 (±0.1)

#	Name	Isotope labeling	Concentration
5	Green Fluorescent Protein	[U-95% 13C; U-95% 15N; U-75% 2H]	1.0 mM
6	sodium chloride		137 mM
7	potassium chloride		2.7 mM
8	sodium phosphate		6.0 mM

Chemical shift validation 3 contents Detail

Properties

Input source #1: Assigned chemical shifts - Assigned chemical shifts	bmr5666_3.str
Diamagnetism of the molecular assembly	True (excluding Oxygen atoms)
Whether the assembly has a disulfide bond	None
Whether the assembly has a other bond	True (see coodinates for details)
Whether the assembly contains a cyclic polymer	None
Overall data processing status	Warning

Disulfide bonds

None

Other bonds (neither disulfide, covalent nor hydrogen bonds, e.g. Zinc–sulphur bond)

Ptnr_site_1	Ptnr_site_2	Redox_state_prediction_1	Redox_state_prediction_2	Distance (Å)
1:65:SER:C	1:66:TYR:N	unknown	unknown	n/a
1:65:SER:C	1:67:GLY:N	unknown	unknown	n/a
1:66:TYR:CA	1:66:TYR:CB	unknown	unknown	n/a

Non-standard residues

None

Content subtype: bmr5666_3.str

#	Content subtype	Saveframe	Status	# of rows	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	shift_set_1	Warning	986		93.0 (chain: 1, length: 229)
1	Chemical shift references	chemical_shift_reference	OK	3		No information

Assigned chemical shifts

Saveframe: shift_set_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: 1, Chain length: 229, Sequence coverage: 93.0%
- Total number of assignments
  - ¹³C chemical shifts: 588
  - ¹⁵N chemical shifts: 199
  - ¹H chemical shifts: 199
- Completeness of assignments
  - Entity_assemble_ID: 1
- Redox state of CYS
- Peptide bond of PRO
- Histogram of normalized assigned chemical shifts
- Random coil index and derived parameters (S² and NMR RMSD)
  - Chain_ID: 1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	1044	588	56.3
¹⁵N chemical shifts	247	199	80.6
¹H chemical shifts	1387	199	14.3

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	458	403	88.0
¹⁵N chemical shifts	219	198	90.4
¹H chemical shifts	470	198	42.1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	586	185	31.6
¹⁵N chemical shifts	28	1	3.6
¹H chemical shifts	917	1	0.1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	119	9	7.6
¹H chemical shifts	119	0	0.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	123	0	0.0
¹⁵N chemical shifts	1	1	100.0
¹H chemical shifts	124	1	0.8

Keywords GFP, NMR, assignment, chromophore

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Found 1 Document (1.11 seconds)

BMRB: 5666

Sample #1

Sample #2

Chemical shift reference

Chemical shift reference

Chemical shift reference

Related entities 1. truncated GFPuv, : 1 : 1 : 106 entities Detail

Interaction partners 1. truncated GFPuv, : 1 interactors Detail

Experiments performed 5 experiments Detail

Instrument

Sample #1

Sample #2

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Chemical shift validation 3 contents Detail