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 50166

Title: P2MIN   PubMed: 32126231

Deposition date: 2020-01-22 Original release date: 2020-05-30

Authors: Orriss, George; To, Vu; Moya-Torres, Aniel; Seabrook, Genevieve; O'Neil, Joe; Stetefeld, Jorg

Citation: Orriss, George; To, Vu; Moya-Torres, Aniel; Seabrook, Genevieve; O'Neil, Joe; Stetefeld, Jorg. "Solution Structure of the Cytoplasmic Domain of NhaP2 a K +/H + Antiporter From Vibrio Cholera"  Biochim. Biophys. Acta Biomembr. 1862, 183225-183225 (2020).

Assembly members:
Cytoplasmic tail of NhaP2 a K+/H+ antiporter from Vibrio cholera, polymer, 165 residues, Formula weight is not available

Natural source:   Common Name: Vibrio cholera   Taxonomy ID: 666   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Vibrio cholera

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

Entity Sequences (FASTA):
Cytoplasmic tail of NhaP2 a K+/H+ antiporter from Vibrio cholera: MTSEWELFIYKLKADKWCIG EPLRNLFMPEGTRIAAVFRD NQLLHPSGSTELCEGDTLCV MAQERDLESLSRLFSEAPEK ASLARFFGDFFLDIEAKLQD VALLYGLDLGELEADAKLKD LVLEHLGETPVLGDYFEWHG LQWVVADVVDWKVTKIGLRE NLYFQ

Data typeCount
13C chemical shifts502
15N chemical shifts137
1H chemical shifts751

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1P2MIN1

Entities:

Entity 1, P2MIN 165 residues - Formula weight is not available

Sequence corresponds to the 158 amino acid fragment consisting of T397 to R555 (the full length NhaP2 protein is 581 amino acid residues) plus an additional 6 residues (ENLYFQ) at the C-terminal end of the molecule which are the remnants of the TEV protease cleavage site after TEV treatment to remove the His6-tag. The 165th and final residue is the initiating Methionine that is required for the bacterial over-expression of the protein and which preceeds the first residue T2 observed in the NMR structure. The 158 amino acid fragment corresponds to the predicted RCK and TrkA-like domains found in the cytoplasmic tail of the NhaP2 K+/H+ antiporter.

1   METTHRSERGLUTRPGLULEUPHEILETYR
2   LYSLEULYSALAASPLYSTRPCYSILEGLY
3   GLUPROLEUARGASNLEUPHEMETPROGLU
4   GLYTHRARGILEALAALAVALPHEARGASP
5   ASNGLNLEULEUHISPROSERGLYSERTHR
6   GLULEUCYSGLUGLYASPTHRLEUCYSVAL
7   METALAGLNGLUARGASPLEUGLUSERLEU
8   SERARGLEUPHESERGLUALAPROGLULYS
9   ALASERLEUALAARGPHEPHEGLYASPPHE
10   PHELEUASPILEGLUALALYSLEUGLNASP
11   VALALALEULEUTYRGLYLEUASPLEUGLY
12   GLULEUGLUALAASPALALYSLEULYSASP
13   LEUVALLEUGLUHISLEUGLYGLUTHRPRO
14   VALLEUGLYASPTYRPHEGLUTRPHISGLY
15   LEUGLNTRPVALVALALAASPVALVALASP
16   TRPLYSVALTHRLYSILEGLYLEUARGGLU
17   ASNLEUTYRPHEGLN

Samples:

sample_1_P2MIN: P2MIN, [U-100% 15N], 550-600 uM; MES-NaOH 20 mM; NaCl 150 mM; TCEP 5 mM; AEBSF (4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride) 0.1 mM; sodium azide 0.02 %v

sample_2_P2MIN: P2MIN, [U-100% 13C; U-100% 15N], 550-600 uM; MES-NaOH 20 mM; NaCl 150 mM; TCEP 5 mM; AEBSF (4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride) 0.1 mM; sodium azide 0.02 %v

sample_3_P2MIN: P2MIN, [U-100% 13C], 550-600 uM; MES-NaOH 20 mM; NaCl 150 mM; TCEP 5 mM; AEBSF (4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride) 0.1 mM; sodium azide 0.02 %v/v

sample_conditions_1_P2MIN_15N_labelled: ionic strength: 0.15 M; pH: 6.5; pressure: 1 atm; temperature: 300 K

sample_conditions_4_P2MIN_15N_and_13C_labelled: ionic strength: 0.15 M; pH: 6.5; pressure: 1 atm; temperature: 300 K

sample_conditions_3_P2MIN_13C_labelled: ionic strength: 0.15 M; pH: 6.5; pressure: 1 atm; temperature: 300 K

sample_conditions_2_P2MIN_13C_labelled: ionic strength: 0.15 M; pH: 6.5; pressure: 1 atm; temperature: 300 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D HNCOsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D HN(CO)CAsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D HNCAsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D CBCA(CO)NHsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D HNCACBsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D HNHAsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D HNHBsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
3D HCCH-TOCSYsample_1_P2MINisotropicsample_conditions_4_P2MIN_15N_and_13C_labelled
3D CC(CO)NHsample_1_P2MINisotropicsample_conditions_4_P2MIN_15N_and_13C_labelled
3D 1H-15N NOESYsample_1_P2MINisotropicsample_conditions_1_P2MIN_15N_labelled
2D 1H-13C HSQCsample_1_P2MINisotropicsample_conditions_3_P2MIN_13C_labelled
3D 1H-13C NOESYsample_1_P2MINisotropicsample_conditions_3_P2MIN_13C_labelled

Software:

NMRPipe, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - processing

SPARKY v3.1.34, Wonghee Lee - chemical shift assignment

TALOS, Cornilescu, Delaglio and Bax - Backbone and sidechain Torsion Angles prediction

PONDEROSA, PONDEROSA - structure solution

TOPSPIN v4.0, Bruker Biospin - data acquisition and analysis

VNMRj, Varian - collection

Relax v4.1, d'Auvergne, E. J. and Gooley, P. R. - data analysis

NMR spectrometers:

  • Bruker Avance 800 MHz
  • Bruker Avance 700 MHz
  • Varian INOVA 600 MHz

Related Database Links:

EXPASY A5F4U3

Download simulated HSQC data in one of the following formats:
CSV: Backbone or all simulated shifts
SPARKY: Backbone or all simulated shifts