BMRB Entry 26851

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR26851

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Title:
1H, 15N, 13C backbone resonance assignments of human soluble catechol O-methyltransferase in complex with 3,5-dinitrocatechol and Sinefungin
Deposition date:
2016-07-13
Original release date:
2019-05-22
Authors:
Baxter, Nicola; Czarnota, Sylwia
Citation:

Citation: Czarnota, Sylwia; Johannissen, Linus; Baxter, Nicola; Rummel, Felix; Wilson, Alex; Cliff, Matthew; Levy, Colin; Scrutton, Nigel; Waltho, Jonathan; Hay, Sam. "Equatorial Active Site Compaction and Electrostatic Reorganization in Catechol-"  ACS Catal. 9, 4394-4401 (2019).
PubMed: 31080692

Assembly members:

Assembly members:
COMT, polymer, 233 residues, 26066 Da.
SINEFUNGIN, non-polymer, 381.387 Da.
3,5-DINITROCATECHOL, non-polymer, 200.106 Da.
MAGNESIUM ION, non-polymer, 24.305 Da.

Natural source:

Natural source:   Common Name: Human   Taxonomy ID: 9606   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Homo sapiens

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pET21a

Entity Sequences (FASTA):

Entity Sequences (FASTA):
COMT: MHHHHHHENLYFQGDTKEQR ILNHVLQHAEPGNAQSVLEA IDTYCEQKEWAMNVGDKKGK IVDAVIQEHQPSVLLELGAY CGYSAVRMARLLSPGARLIT IEINPDCAAITQRMVDFAGV KDKVTLVVGASQDIIPQLKK KYDVDTLDMVFLDHWKDRYL PDTLLLEECGLLRKGTVLLA DNVICPGAPDFLAHVRGSSC FECTHYQSFLEYREVVDGLE KAIYKGPGSEAGP

Data sets:
Data typeCount
13C chemical shifts639
15N chemical shifts204
1H chemical shifts204

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1COMT1
2Sinefungin2
3DNC3
4Mg4

Entities:

Entity 1, COMT 233 residues - 26066 Da.

The human S-COMT construct used in this study has a 13 residue His-tag and cloning sequence positioned at the N-terminus (MHHHHHHENLYFQG ), where G14 in this construct corresponds with G2 in the canonical S-COMT sequence. This construct also contains valine (rather than methionine) at the allelic polymorphism position located at residue 108 of S-COMT.

1   METHISHISHISHISHISHISGLUASNLEU
2   TYRPHEGLNGLYASPTHRLYSGLUGLNARG
3   ILELEUASNHISVALLEUGLNHISALAGLU
4   PROGLYASNALAGLNSERVALLEUGLUALA
5   ILEASPTHRTYRCYSGLUGLNLYSGLUTRP
6   ALAMETASNVALGLYASPLYSLYSGLYLYS
7   ILEVALASPALAVALILEGLNGLUHISGLN
8   PROSERVALLEULEUGLULEUGLYALATYR
9   CYSGLYTYRSERALAVALARGMETALAARG
10   LEULEUSERPROGLYALAARGLEUILETHR
11   ILEGLUILEASNPROASPCYSALAALAILE
12   THRGLNARGMETVALASPPHEALAGLYVAL
13   LYSASPLYSVALTHRLEUVALVALGLYALA
14   SERGLNASPILEILEPROGLNLEULYSLYS
15   LYSTYRASPVALASPTHRLEUASPMETVAL
16   PHELEUASPHISTRPLYSASPARGTYRLEU
17   PROASPTHRLEULEULEUGLUGLUCYSGLY
18   LEULEUARGLYSGLYTHRVALLEULEUALA
19   ASPASNVALILECYSPROGLYALAPROASP
20   PHELEUALAHISVALARGGLYSERSERCYS
21   PHEGLUCYSTHRHISTYRGLNSERPHELEU
22   GLUTYRARGGLUVALVALASPGLYLEUGLU
23   LYSALAILETYRLYSGLYPROGLYSERGLU
24   ALAGLYPRO

Entity 2, Sinefungin - 381.387 Da.

1   SFG

Entity 3, DNC - 200.106 Da.

1   DNC

Entity 4, Mg - 24.305 Da.

1   MG

Samples:

sample_1: COMT, [U-99% 13C; U-99% 15N], 0.5 mM; Sinefungin 5 mM; DNC 5 mM; MG 2.5 mM; DTT 10 mM; sodium chloride 50 mM; TRIS 50 mM; H2O 90%; D2O 10%

sample_conditions_1: ionic strength: 100 mM; pH: 7.5; pressure: 1 atm; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
3D HNCOsample_1isotropicsample_conditions_1
3D HNCAsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D HN(CO)CACBsample_1isotropicsample_conditions_1
3D HN(CA)COsample_1isotropicsample_conditions_1
3D 1H-15N TROSYsample_1isotropicsample_conditions_1
3D HN(CO)CAsample_1isotropicsample_conditions_1

Software:

TOPSPIN v3.2, Bruker Biospin - collection, processing

Felix, Accelrys Software Inc. - chemical shift assignment, peak picking

CCPN_Analysis v2.4, CCPN - chemical shift assignment, peak picking

NMR spectrometers:

  • Bruker Avance 800 MHz

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