BMRBj - BMREntryMaster

Found 1 Document (0.461 seconds)

BMRB: 30888

7M2M

NMR Structure of GCAP5

Authors

Ames, J.B., Cudia, D.L.

Assembly

Guanylate cyclase activator 1A

Entity

1. Guanylate cyclase activator 1A, entity 1 (polymer), 198 monomers, 22471.20 Da Detail

XGDSSSMSAT ELSACKCHQW YRKFMTECPS GQLTFYEFKK FFGLKNLSEK SNAYVNTMFK TFDIDDDGCI DFMEYVAALS LVLKGGVQQK LRWYFKLFDM DGSGCIDKDE LLLIFKAVQA INGAEPEISA EDLADIVFNK IDVNGDGELS LEEFMEGISA DEKISEMLTQ SLDLTRIVSN IYNDSYIEQE AEIIEDQA

2. Guanylate cyclase activator 1A, entity MG (non-polymer), 24.305 Da

Total weight

22495.504 Da

Max. entity weight

22471.2 Da

Source organism

Danio rerio

Exptl. method

solution NMR

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 91.4 %, Completeness: 65.3 %, Completeness (bb): 85.6 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	65.3 % (1505 of 2304)	60.6 % (720 of 1189)	68.7 % (618 of 899)	77.3 % (167 of 216)
Backbone	85.6 % (1008 of 1178)	83.2 % (336 of 404)	87.6 % (507 of 579)	84.6 % (165 of 195)
Sidechain	50.6 % (664 of 1311)	48.9 % (384 of 785)	54.9 % (277 of 505)	14.3 % (3 of 21)
Aromatic	17.3 % (37 of 214)	15.9 % (17 of 107)	17.1 % (18 of 105)	100.0 % (2 of 2)
Methyl	77.5 % (158 of 204)	78.4 % (80 of 102)	76.5 % (78 of 102)

1. entity 1

Show sample condition

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 6.0, Details 0.5 mM [U-100% 15N] GCAP5, 2.0 mM MAGNESIUM ION, 10 mM [U-100% 2H] MES, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
1	GCAP5	[U-100% 15N]	0.5 mM
2	MAGNESIUM ION	natural abundance	2.0 mM
3	MES	[U-100% 2H]	10 mM

Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 6.0, Details 0.5 mM [U-100% 13C; U-100% 15N] GCAP5, 2.0 mM MAGNESIUM ION, 10 mM [U-100% 2H] MES, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
4	GCAP5	[U-100% 13C; U-100% 15N]	0.5 mM
5	MAGNESIUM ION	natural abundance	2.0 mM
6	MES	[U-100% 2H]	10 mM

Release date

2021-03-17

Citation

NMR and EPR-DEER Structure of a Dimeric Guanylate Cyclase Activator Protein-5 from Zebrafish Photoreceptors
Cudia, D., Roseman, G.P., Assafa, T.E., Shahu, M.K., Scholten, A., Menke-Sell, S.K., Yamada, H., Koch, K.W., Milhauser, G., Ames, J.B.
Biochemistry (2021), 60, 3058-3070, PubMed 34609135 , DOI 10.1021/acs.biochem.1c00612 , Abstract

Related entities 1. Guanylate cyclase activator 1A, entity 1, : 1 : 184 entities Detail

Experiments performed 6 experiments Detail

1 2D 1H-15N HSQC

Instrument

Bruker AVANCE III - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 6.0, Details 0.5 mM [U-100% 15N] GCAP5, 2.0 mM MAGNESIUM ION, 10 mM [U-100% 2H] MES, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
1	GCAP5	[U-100% 15N]	0.5 mM
2	MAGNESIUM ION	natural abundance	2.0 mM
3	MES	[U-100% 2H]	10 mM

2 2D 1H-13C HSQC aliphatic

Instrument

Bruker AVANCE III - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 6.0, Details 0.5 mM [U-100% 13C; U-100% 15N] GCAP5, 2.0 mM MAGNESIUM ION, 10 mM [U-100% 2H] MES, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
4	GCAP5	[U-100% 13C; U-100% 15N]	0.5 mM
5	MAGNESIUM ION	natural abundance	2.0 mM
6	MES	[U-100% 2H]	10 mM

3 3D HNCACB

Instrument

Bruker AVANCE III - 800 MHz

Sample

#	Name	Isotope labeling	Concentration
4	GCAP5	[U-100% 13C; U-100% 15N]	0.5 mM
5	MAGNESIUM ION	natural abundance	2.0 mM
6	MES	[U-100% 2H]	10 mM

4 3D CBCA(CO)NH

Instrument

Bruker AVANCE III - 800 MHz

Sample

#	Name	Isotope labeling	Concentration
4	GCAP5	[U-100% 13C; U-100% 15N]	0.5 mM
5	MAGNESIUM ION	natural abundance	2.0 mM
6	MES	[U-100% 2H]	10 mM

5 3D HCCH-TOCSY

Instrument

Bruker AVANCE III - 800 MHz

Sample

#	Name	Isotope labeling	Concentration
4	GCAP5	[U-100% 13C; U-100% 15N]	0.5 mM
5	MAGNESIUM ION	natural abundance	2.0 mM
6	MES	[U-100% 2H]	10 mM

6 3D 1H-13C NOESY aliphatic

Instrument

Bruker AVANCE III - 800 MHz

Sample

#	Name	Isotope labeling	Concentration
4	GCAP5	[U-100% 13C; U-100% 15N]	0.5 mM
5	MAGNESIUM ION	natural abundance	2.0 mM
6	MES	[U-100% 2H]	10 mM

Chemical shift validation 4 contents Detail

Properties

Input source #1: Assigned chemical shifts - Assigned chemical shifts	bmr30888_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	None
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)

None

Non-standard residues

None

Content subtype: bmr30888_3.str

#	Content subtype	Saveframe	Status	# of rows	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chemical_shifts_1	Warning	1551		88.4 (chain: 1, length: 198)
1	Chemical shift references	chem_shift_reference_1	OK	3		No information

Assigned chemical shifts

Saveframe: assigned_chemical_shifts_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: 1, Chain length: 198, Sequence coverage: 88.4%
- Total number of assignments
  - ¹H chemical shifts: 812
  - ¹³C chemical shifts: 579
  - ¹⁵N chemical shifts: 160
- Completeness of assignments
  - Entity_assemble_ID: 1
- Redox state of CYS
- Peptide bond of PRO
- Rotameric state of ILE , LEU, and VAL
- 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 (%)
¹H chemical shifts	1214	654	53.9
¹³C chemical shifts	913	579	63.4
¹⁵N chemical shifts	216	160	74.1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	406	317	78.1
¹³C chemical shifts	400	331	82.8
¹⁵N chemical shifts	195	158	81.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	808	337	41.7
¹³C chemical shifts	513	248	48.3
¹⁵N chemical shifts	21	2	9.5

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	110	79	71.8
¹³C chemical shifts	110	80	72.7

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	107	2	1.9
¹³C chemical shifts	105	0	0.0
¹⁵N chemical shifts	2	2	100.0

Keywords Ca2+, EF-hand, GCAP5, METAL BINDING PROTEIN, Mg2+, guanylate cyclase, phototransduction, vision

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Found 1 Document (0.461 seconds)

BMRB: 30888

Sample #1

Sample #2

Related entities 1. Guanylate cyclase activator 1A, entity 1, : 1 : 184 entities Detail

Experiments performed 6 experiments Detail

Instrument

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Chemical shift validation 4 contents Detail