BMRB Entry 30819

Title:
NMR Structure of a tRNA 2'-phosphotransferase from Runella slithyformis
Deposition date:
2020-11-30
Original release date:
2021-10-08
Authors:
Alphonse, S.; Dantuluri, S.; Banerjee, A.; Shuman, S.; Ghose, R.
Citation:

Citation: Alphonse, Sebastien; Banerjee, Ankan; Dantuluri, Swathi; Shuman, Stewart; Ghose, Ranajeet. "NMR solution structures of Runella slithyformis RNA 2'-phosphotransferase Tpt1 provide insights into NAD+ binding and specificity"  Nucleic Acids Res. 49, 9607-9624 (2021).
PubMed: 33880546

Assembly members:

Assembly members:
entity_1, polymer, 178 residues, 19846.059 Da.

Natural source:

Natural source:   Common Name: Runella slithyformis   Taxonomy ID: 106   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Runella slithyformis

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli BL21(DE3)

Data sets:
Data typeCount
13C chemical shifts718
15N chemical shifts179
1H chemical shifts1139

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
  • Hide all

Assembly:

Entity Assembly IDEntity NameEntity ID
1unit_11

Entities:

Entity 1, unit_1 178 residues - 19846.059 Da.

1   GLYSERHISMETVALLYSVALSERLYSPHE
2   LEUSERLEUVALLEUARGHISASNPROALA
3   LEUILEGLYLEUASPLEUASPALAASNGLY
4   TRPALAPROVALLYSGLULEULEUALALYS
5   METLYSALALYSGLYHISGLYILESERMET
6   GLUGLULEULYSHISILEVALGLUTHRASN
7   SERLYSLYSARGPHEALAPHESERGLUASN
8   PHEGLULYSILEARGALAASNGLNGLYHIS
9   SERVALGLUVALASPLEUGLYTYRGLULYS
10   GLNVALPROPROALAVALLEUPHEHISGLY
11   THRALAGLULYSASNPHEASPLEUILELEU
12   LYSASPGLYILELYSLYSMETSERARGHIS
13   HISVALHISLEUSERGLNASPILETHRTHR
14   ALAARGLYSVALGLYMETARGHISGLYLYS
15   PROVALVALLEUSERVALASPALALYSGLY
16   METALAASPGLYGLYPHEASPPHETYRLEU
17   SERASNASNGLYVALTRPLEUILEASPPHE
18   VALPROALAGLUPHEILELYSVAL

Samples:

sample_1: Tpt1, [U-13C; U-15N], 300 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.04 % v/v; DSS 50 uM

sample_2: Tpt1, [U-13C; U-15N], 360 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.04 % v/v; DSS 50 uM

sample_3: Tpt1, [U-13C; U-15N], 300 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 1 mM; sodium azide 0.05 % w/v; DSS 50 uM

sample_4: Tpt1, [U-13C; U-15N], 400 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.05 % w/v; DSS 50 mM

sample_5: Tpt1, [U-2H; U-15N]; selective methyl groups, 400 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.05 % w/v; DSS 50 uM

sample_6: Tpt1, [U-2H; U-15N]; selective methyl groups, 350 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.05 % w/v; DSS 50 uM

sample_7: Tpt1, [U-13C; U-15N], 420 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.04 % w/v; DSS 50 mM

sample_8: Tpt1, [U-13C; U-15N], 380 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.04 % w/v; DSS 50 mM

sample_9: Tpt1, [U-13C; U-15N], 400 uM; HEPES 20 mM; sodium chloride 200 mM; DTT 2 mM; EDTA, [U-100% 2H], 2 mM; glycerol, [U-100% 2H], 5 % v/v; AEBSF protease inhibitor 0.5 mM; sodium azide 0.04 % w/v; DSS 50 mM

sample_conditions_1: pH: 7; pressure: 1 atm; temperature: 290.15 K

sample_conditions_2: pH: 7 pH*; pressure: 1 atm; temperature: 290.15 K

sample_conditions_3: pH: 7; pressure: 1 atm; temperature: 290.15 K

sample_conditions_4: pH: 7 pH*; pressure: 1 atm; temperature: 290.15 K

Experiments:

NameSampleSample stateSample conditions
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D 1H-13C NOESY aliphaticsample_2isotropicsample_conditions_1
3D 1H-13C NOESY aromaticsample_3isotropicsample_conditions_1
3D 1H-13C NOESY aromaticsample_4isotropicsample_conditions_1
3D 1H-13C Methyl NOESYsample_5isotropicsample_conditions_2
1H-13C-13C-1H 4D NOESYsample_6isotropicsample_conditions_2
3D HNCAsample_7isotropicsample_conditions_3
3D HNCOsample_8isotropicsample_conditions_3
3D HN(CA)COsample_8isotropicsample_conditions_3
3D CBCA(CO)NHsample_8isotropicsample_conditions_3
3D HNCACBsample_8isotropicsample_conditions_3
3D C(CO)NHsample_8isotropicsample_conditions_3
3D H(CCO)NHsample_8isotropicsample_conditions_3
3D CCH-TOCSYsample_9isotropicsample_conditions_4
3D HCCH-TOCSYsample_9isotropicsample_conditions_4

Software:

NMRPipe v10.3 Revision 2019.220.13.40, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - processing

TopSpin v3.5p15, Bruker Biospin - collection

NMRView, Johnson, One Moon Scientific - chemical shift assignment

ARIA v2.3.2, Linge, O'Donoghue and Nilges - structure calculation

X-PLOR NIH v2.52, Schwieters, Kuszewski, Tjandra and Clore - refinement

NMR spectrometers:

  • Bruker AVANCE III HD 800 MHz
  • Bruker AVANCE III HD 700 MHz
  • Bruker AVANCE III 600 MHz
  • Bruker AVANCE III HD 800 MHz

Download HSQC peak lists in one of the following formats:
CSV: Backbone or all simulated peaks
SPARKY: Backbone or all simulated peaks