BMRB: 34748

Solution NMR structure of full-length Nsp1 from SARS-CoV-2

Authors

Wang, Y., Kirkpatrick, J.P., Carlomagno, T.

Assembly

Host translation inhibitor nsp1

Entity

1. Host translation inhibitor nsp1 (polymer), 182 monomers, 19903.18 Da Detail

GAMESLVPGF NEKTHVQLSL PVLQVRDVLV RGFGDSVEEV LSEARQHLKD GTCGLVEVEK GVLPQLEQPY VFIKRSDART APHGHVMVEL VAELEGIQYG RSGETLGVLV PHVGEIPVAY RKVLLRKNGN KGAGGHSYGA DLKSFDLGDE LGTDPYEDFQ ENWNTKHSSG VTRELMRELN GG

Formula weight

19903.18 Da

Source organism

Severe acute respiratory syndrome coronavirus 2

Exptl. method

solution NMR

Data set

assigned_chemical_shifts, spectral_peak_list

Chem. Shift Complete

Sequence coverage: 99.5 %, Completeness: 98.2 %, Completeness (bb): 98.5 % Detail

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Polymer type: polypeptide(L)

	Total	1H	13C	15N
All	98.2 % (2030 of 2068)	98.5 % (1052 of 1068)	98.6 % (791 of 802)	94.4 % (187 of 198)
Backbone	98.5 % (1060 of 1076)	98.7 % (374 of 379)	98.7 % (516 of 523)	97.7 % (170 of 174)
Sidechain	98.1 % (1129 of 1151)	98.4 % (678 of 689)	99.1 % (434 of 438)	70.8 % (17 of 24)
Aromatic	93.8 % (122 of 130)	93.8 % (61 of 65)	93.8 % (60 of 64)	100.0 % (1 of 1)
Methyl	100.0 % (210 of 210)	100.0 % (105 of 105)	100.0 % (105 of 105)

1. entity 1

GAMESLVPGF NEKTHVQLSL PVLQVRDVLV RGFGDSVEEV LSEARQHLKD GTCGLVEVEK GVLPQLEQPY VFIKRSDART APHGHVMVEL VAELEGIQYG RSGETLGVLV PHVGEIPVAY RKVLLRKNGN KGAGGHSYGA DLKSFDLGDE LGTDPYEDFQ ENWNTKHSSG VTRELMRELN GG

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Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 6.5 (±0.1), Details 600 uM [U-13C; U-15N] Nsp1, 50 mM sodium phosphate, 200 mM sodium chloride, 2 mM DTT, 2 mM EDTA, 0.01 % w/v sodium azide, 10 % v/v [U-2H] D2O, 90% H2O/10% D2O.

#	Name	Isotope labeling	Type	Concentration
1	Nsp1	[U-13C; U-15N]		600 (±100.0) uM
2	sodium phosphate	natural abundance		50 (±5.0) mM
3	sodium chloride	natural abundance		200 (±10.0) mM
4	DTT	natural abundance		2 (±0.1) mM
5	EDTA	natural abundance		2 (±0.1) mM
6	sodium azide	natural abundance		0.01 (±0.001) % w/v
7	D2O	[U-2H]		10 (±0.5) % v/v

Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 6.5 (±0.1), Details 500 uM [U-10% 13C; U-100% 15N] Nsp1, 50 mM sodium phosphate, 200 mM sodium chloride, 2 mM DTT, 2 mM EDTA, 0.01 % w/v sodium azide, 10 % v/v [U-2H] D2O, 90% H2O/10% D2O.

#	Name	Isotope labeling	Type	Concentration
8	Nsp1	[U-10% 13C; U-100% 15N]		500 (±100.0) uM
9	sodium phosphate	natural abundance		50 (±5.0) mM
10	sodium chloride	natural abundance		200 (±10.0) mM
11	DTT	natural abundance		2 (±0.1) mM
12	EDTA	natural abundance		2 (±0.1) mM
13	sodium azide	natural abundance		0.01 (±0.001) % w/v
14	D2O	[U-2H]		10 (±0.5) % v/v

Sample #3

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 6.5 (±0.1), Details 500 uM [U-10% 13C; U-100% 15N] Nsp1, 50 mM sodium phosphate, 200 mM sodium chloride, 2 mM DTT, 2 mM EDTA, 0.01 % w/w sodium azide, 10 % v/v [U-2H] D2O, 12 mg/mL Pf1 phage, 90% H2O/10% D2O.

#	Name	Isotope labeling	Type	Concentration
15	Nsp1	[U-10% 13C; U-100% 15N]		500 (±100.0) uM
16	sodium phosphate	natural abundance		50 (±5.0) mM
17	sodium chloride	natural abundance		200 (±10.0) mM
18	DTT	natural abundance		2 (±0.1) mM
19	EDTA	natural abundance		2 (±0.1) mM
20	sodium azide	natural abundance		0.01 (±0.001) % w/w
21	D2O	[U-2H]		10 (±0.5) % v/v
22	Pf1 phage	natural abundance		12 (±1.0) mg/mL

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Release date

2022-08-22

Citation 1

¹H, ¹³C, and ¹⁵N backbone chemical-shift assignments of SARS-CoV-2 non-structural protein 1 (leader protein)
Wang, Y., Kirkpatrick, J., zur Lage, S., Korn, S., Neissner, K., Schwalbe, H., Schlundt, A., Carlomagno, T.
Biomol. NMR Assign. (2021), 15, 287-295, PubMed 33770349 , DOI 10.1007/s12104-021-10019-6 , Abstract

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Citation 2

Structural insights into the activity regulation of full-length non-structural protein 1 from SARS-CoV-2
Wang, Y., Kirkpatrick, J.P., zur Lage, S., Carlomagno, T.

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Entries sharing articles BMRB: 1 entries Detail

Related entities 1. Host translation inhibitor nsp1, : 1 : 13 : 9 entities Detail

Interaction partners 1. Host translation inhibitor nsp1, : 1 interactors Detail

Experiments performed 31 experiments Detail