BMRB Entry 27073

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR27073

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Title:
Ile- 1 Methyl resonance assignment of the Hepatitis C Virus RNA-dependent RNA-polymerase NS5B (strain JFH-1).
Deposition date:
2017-04-13
Original release date:
2017-09-13
Authors:
Hanoulle, Xavier; Launay, Helene; Mamigonian-Bessa, Luiza; Cantrelle, Francois-Xavier; Schneider, Robert
Citation:

Citation: Bessa, Luiza; Launay, Helene; Dujardin, Marie; Cantrelle, Francois-Xavier; Lippens, Guy; Landrieu, Isabelle; Schneider, Robert; Hanoulle, Xavier. "NMR reveals the intrinsically disordered domain 2 of NS5A protein as an allosteric regulator of the hepatitis C virus RNA polymerase NS5B"  J. Biol. Chem. 117, 813766-813766 (2017).
PubMed: 28912275

Assembly members:

Assembly members:
HCV_polymerase_polypeptide, polymer, 577 residues, Formula weight is not available

Natural source:

Natural source:   Common Name: Hepatitis C virus   Taxonomy ID: 11103   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Hepacivirus NCV

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pET21

Entity Sequences (FASTA):

Entity Sequences (FASTA):
HCV_polymerase_polypeptide: SMSYSWTGALITPCSPEEEK LPINPLSNSLLRYHNKVYCT TSKSASQRAKKVTFDRTQVL DAHYDSVLKDIKLAASKVSA RLLTLEEACQLTPPHSARSK YGFGAKEVRSLSGRAVNHIK SVWKDLLEDPQTPIPTTIMA KNEVFCVDPAKGGKKPARLI VYPDLGVRVCEKMALYDITQ KLPQAVMGASYGFQYSPAQR VEYLLKAWAEKKDPMGFSYD TRCFDSTVTERDIRTEESIY QACSLPEEARTAIHSLTERL YVGGPMFNSKGQTCGYRRCR ASGVLTTSMGNTITCYVKAL AACKAAGIVAPTMLVCGDDL VVISESQGTEEDERNLRAFT EAMTRYSAPPGDPPRPEYDL ELITSCSSNVSVALGPRGRR RYYLTRDPTTPLARAAWETV RHSPINSWLGNIIQYAPTIW VRMVLMTHFFSILMVQDTLD QNLNFEMYGSVYSVNPLDLP AIIERLHGLDAFSMHTYSHH ELTRVASALRKLGAPPLRVW KSRARAVRASLISRGGKAAV CGRYLFNWAVKTKLKLTPLP EARLLDLSSWFTVGAGGGDI FHSVSRARPRSHHHHHH

Data sets:
Data typeCount
13C chemical shifts20
1H chemical shifts60

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
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Assembly:

Entity Assembly IDEntity NameEntity ID
1NS5Bd211

Entities:

Entity 1, NS5Bd21 577 residues - Formula weight is not available

This is the soluble HCV polymerase without its 21 C-terminal residues (delta21). Residues 571-577 correspond to a non-native affinity tag.

1   SERMETSERTYRSERTRPTHRGLYALALEU
2   ILETHRPROCYSSERPROGLUGLUGLULYS
3   LEUPROILEASNPROLEUSERASNSERLEU
4   LEUARGTYRHISASNLYSVALTYRCYSTHR
5   THRSERLYSSERALASERGLNARGALALYS
6   LYSVALTHRPHEASPARGTHRGLNVALLEU
7   ASPALAHISTYRASPSERVALLEULYSASP
8   ILELYSLEUALAALASERLYSVALSERALA
9   ARGLEULEUTHRLEUGLUGLUALACYSGLN
10   LEUTHRPROPROHISSERALAARGSERLYS
11   TYRGLYPHEGLYALALYSGLUVALARGSER
12   LEUSERGLYARGALAVALASNHISILELYS
13   SERVALTRPLYSASPLEULEUGLUASPPRO
14   GLNTHRPROILEPROTHRTHRILEMETALA
15   LYSASNGLUVALPHECYSVALASPPROALA
16   LYSGLYGLYLYSLYSPROALAARGLEUILE
17   VALTYRPROASPLEUGLYVALARGVALCYS
18   GLULYSMETALALEUTYRASPILETHRGLN
19   LYSLEUPROGLNALAVALMETGLYALASER
20   TYRGLYPHEGLNTYRSERPROALAGLNARG
21   VALGLUTYRLEULEULYSALATRPALAGLU
22   LYSLYSASPPROMETGLYPHESERTYRASP
23   THRARGCYSPHEASPSERTHRVALTHRGLU
24   ARGASPILEARGTHRGLUGLUSERILETYR
25   GLNALACYSSERLEUPROGLUGLUALAARG
26   THRALAILEHISSERLEUTHRGLUARGLEU
27   TYRVALGLYGLYPROMETPHEASNSERLYS
28   GLYGLNTHRCYSGLYTYRARGARGCYSARG
29   ALASERGLYVALLEUTHRTHRSERMETGLY
30   ASNTHRILETHRCYSTYRVALLYSALALEU
31   ALAALACYSLYSALAALAGLYILEVALALA
32   PROTHRMETLEUVALCYSGLYASPASPLEU
33   VALVALILESERGLUSERGLNGLYTHRGLU
34   GLUASPGLUARGASNLEUARGALAPHETHR
35   GLUALAMETTHRARGTYRSERALAPROPRO
36   GLYASPPROPROARGPROGLUTYRASPLEU
37   GLULEUILETHRSERCYSSERSERASNVAL
38   SERVALALALEUGLYPROARGGLYARGARG
39   ARGTYRTYRLEUTHRARGASPPROTHRTHR
40   PROLEUALAARGALAALATRPGLUTHRVAL
41   ARGHISSERPROILEASNSERTRPLEUGLY
42   ASNILEILEGLNTYRALAPROTHRILETRP
43   VALARGMETVALLEUMETTHRHISPHEPHE
44   SERILELEUMETVALGLNASPTHRLEUASP
45   GLNASNLEUASNPHEGLUMETTYRGLYSER
46   VALTYRSERVALASNPROLEUASPLEUPRO
47   ALAILEILEGLUARGLEUHISGLYLEUASP
48   ALAPHESERMETHISTHRTYRSERHISHIS
49   GLULEUTHRARGVALALASERALALEUARG
50   LYSLEUGLYALAPROPROLEUARGVALTRP
51   LYSSERARGALAARGALAVALARGALASER
52   LEUILESERARGGLYGLYLYSALAALAVAL
53   CYSGLYARGTYRLEUPHEASNTRPALAVAL
54   LYSTHRLYSLEULYSLEUTHRPROLEUPRO
55   GLUALAARGLEULEUASPLEUSERSERTRP
56   PHETHRVALGLYALAGLYGLYGLYASPILE
57   PHEHISSERVALSERARGALAARGPROARG
58   SERHISHISHISHISHISHIS

Samples:

sample_1: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_2: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_3: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_4: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_5: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_6: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_7: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_8: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_9: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_14: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_10: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_11: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_12: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; potassium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_13: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_15: HCV polymerase polypeptide, [Ile-Cd1-13CH3; U-2H], 70 uM; sodium phosphate 300 mM; sodium chloride 50 mM; magnesium chloride 2 mM; EDTA 1 mM; THP 4 mM; TMSP 0.150 mM

sample_conditions_1: ionic strength: 350 mM; pH: 6.8; pressure: 1 atm; temperature: 305 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-13C HMQCsample_1isotropicsample_conditions_1
3D 1H-13C-1H NOESYsample_1isotropicsample_conditions_1
3D 1H-13C-13C NOESYsample_1isotropicsample_conditions_1
2D 1H-13C HMQCsample_2isotropicsample_conditions_1
2D 1H-13C HMQCsample_3isotropicsample_conditions_1
2D 1H-13C HMQCsample_4isotropicsample_conditions_1
2D 1H-13C HMQCsample_5isotropicsample_conditions_1
2D 1H-13C HMQCsample_6isotropicsample_conditions_1
2D 1H-13C HMQCsample_7isotropicsample_conditions_1
2D 1H-13C HMQCsample_8isotropicsample_conditions_1
2D 1H-13C HMQCsample_9isotropicsample_conditions_1
2D 1H-13C HMQCsample_10isotropicsample_conditions_1
2D 1H-13C HMQCsample_11isotropicsample_conditions_1
2D 1H-13C HMQCsample_12isotropicsample_conditions_1
2D 1H-13C HMQCsample_13isotropicsample_conditions_1
2D 1H-13C HMQCsample_14isotropicsample_conditions_1
2D 1H-13C HMQCsample_15isotropicsample_conditions_1

Software:

TOPSPIN v3.5, Bruker Biospin - collection, processing

SPARKY vNMRFAM-SPARKY, Goddard - chemical shift assignment, data analysis

NMR spectrometers:

  • Bruker Avance 900 MHz

Related Database Links:

UNP Q99IB8
AlphaFold Q99IB8