BMRB

Biological Magnetic Resonance Data Bank


A Repository for Data from NMR Spectroscopy on Proteins, Peptides, Nucleic Acids, and other Biomolecules
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BMRB Entry 16486

Title: Solution NMR structure of the ACT domain from GTP pyrophosphokinase of Chlorobium tepidum. Northeast Structural Genomics Consortium Target CtR148A

Authors: Eletsky, Alexander; Garcia, Erwin; Wang, Huang; Ciccosanti, Colleen; Jiang, Mei; Nair, Rajesh; Rost, Burkhard; Acton, Thomas; Xiao, Rong; Everett, John; Lee, Hsiau-Wei; Prestegard, James; Montelione, Gaetano; Szyperski, Thomas

Citation: Eletsky, Alexander; Garcia, Erwin; Wang, Huang; Ciccosanti, Colleen; Jiang, Mei; Nair, Rajesh; Rost, Burkhard; Acton, Thomas; Xiao, Rong; Everett, John; Lee, Hsiau-Wei; Prestegard, James; Montelione, Gaetano; Szyperski, Thomas. "Solution NMR structure of the ACT domain from GTP pyrophosphokinase of Chlorobium tepidum."  Proteins: Struct. Funct. Genet. ., .-..

Assembly members:
CtR148A, polymer, 88 residues, 10050.751 Da.

Natural source:   Common Name: Chlorobium tepidum   Taxonomy ID: 194439   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Chlorobium tepidum

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

Entity Sequences (FASTA):
CtR148A: MTDFLAGIRIVGEDKNGMTN QITGVISKFDTNIRTIVLNA KDGIFTCNLMIFVKNTDKLT TLMDKLRKVQGVFTVERLSN LEHHHHHH

Data sets:
Data typeCount
13C chemical shifts368
15N chemical shifts94
1H chemical shifts630

Additional metadata:

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

Entity Assembly IDEntity NameEntity ID
1entity_11
2entity_21

Entities:

Entity 1, entity_1 88 residues - 10050.751 Da.

Residues 2-80 correspond to the 653-731 fragment of the native protein. Residues 81-88 represent a non-native affinity tag. Residue M1 is a result of a new start codon.

1   METTHRASPPHELEUALAGLYILEARGILE
2   VALGLYGLUASPLYSASNGLYMETTHRASN
3   GLNILETHRGLYVALILESERLYSPHEASP
4   THRASNILEARGTHRILEVALLEUASNALA
5   LYSASPGLYILEPHETHRCYSASNLEUMET
6   ILEPHEVALLYSASNTHRASPLYSLEUTHR
7   THRLEUMETASPLYSLEUARGLYSVALGLN
8   GLYVALPHETHRVALGLUARGLEUSERASN
9   LEUGLUHISHISHISHISHISHIS

Samples:

NC: CtR148A, [U-100% 13C; U-100% 15N], 0.3 mM; MES 20 mM; sodium chloride 200 mM; calcium chloride 5 mM; DTT 10 mM; DSS 50 uM; sodium azide 0.05%; D2O 10%; H2O 90%

NC5: CtR148A, [U-5% 13C; U-100% 15N], 0.5 mM; MES 20 mM; sodium chloride 200 mM; calcium chloride 5 mM; DTT 10 mM; DSS 50 uM; sodium azide 0.05%; D2O 10%; H2O 90%

MIX: CtR148A, [U-100% 13C; U-100% 15N], 0.5 mM; MES 20 mM; sodium chloride 200 mM; calcium chloride 5 mM; DTT 10 mM; DSS 50 uM; sodium azide 0.05%; CtR148A 0.5 mM; D2O 10%; H2O 90%

NC5_PEG: CtR148A, [U-5% 13C; U-100% 15N], 0.3 mM; MES 18 mM; sodium chloride 180 mM; calcium chloride 4.5 mM; DTT 9 mM; DSS 45 uM; sodium azide 0.045%; polyethylene glycol 4%; D2O 16%; H2O 84%

NC5_PAAG: CtR148A, [U-5% 13C; U-100% 15N], 0.45 mM; MES 18 mM; sodium chloride 180 mM; calcium chloride 4.5 mM; DTT 9 mM; DSS 45 uM; sodium azide 0.045%; polyacrylamide 7%; D2O 14%; H2O 86%

sample_conditions_1: ionic strength: 225 mM; pH: 6.5; pressure: 1 atm; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
1D 15N T1NCisotropicsample_conditions_1
1D 15N T2NCisotropicsample_conditions_1
2D 1H-15N HSQCNCisotropicsample_conditions_1
2D 1H-13C HSQC aliphaticNCisotropicsample_conditions_1
2D 1H-13C CT-HSQC aliphaticNCisotropicsample_conditions_1
2D 1H-13C CT-HSQC aromaticNCisotropicsample_conditions_1
3D CBCA(CO)NHNCisotropicsample_conditions_1
3D HNCACBNCisotropicsample_conditions_1
3D HNCONCisotropicsample_conditions_1
3D HN(CA)CONCisotropicsample_conditions_1
3D HBHA(CO)NHNCisotropicsample_conditions_1
3D HCCH-COSYNCisotropicsample_conditions_1
3D HCCH-TOCSYNCisotropicsample_conditions_1
3D 1H-13C NOESYNCisotropicsample_conditions_1
3D 1H-15N/13C NOESYNCisotropicsample_conditions_1
2D 1H-13C CT-HSQC methylNC5isotropicsample_conditions_1
2D 1H-15N HSQC J-modulatedNC5isotropicsample_conditions_1
2D 1H-15N HSQC J-modulatedNC5_PEGanisotropicsample_conditions_1
2D 1H-15N HSQC J-modulatedNC5_PAAGanisotropicsample_conditions_1
3D 1H-13C X-filtered NOESYMIXisotropicsample_conditions_1

Software:

VNMRJ v2.1B, Varian - collection

PROSA v6.4, Guntert - processing

CARA v1.8.4, Keller and Wuthrich - chemical shift assignment, data analysis, peak picking

PINE v1.0, Bahrami, Markley, Assadi, and Eghbalnia - chemical shift assignment

TALOS+ v1.2009.0721.18, Yang,Cornilescu, Delaglio and Bax - data analysis

CYANA v3.0, Guntert, Mumenthaler and Wuthrich - structure solution

CNS v1.2.1, Brunger, Adams, Clore, Gros, Nilges and Read - refinement

AutoStruct v2.2.1, Huang, Tejero, Powers and Montelione - structure validation

PSVS v1.3, Bhattacharya and Montelione - structure validation

NMR spectrometers:

  • Varian INOVA 750 MHz
  • Varian INOVA 600 MHz
  • Varian INOVA 600 MHz

Related Database Links:

PDB
GB AAM72771
REF NP_662429 WP_010933210