BMRB Entry 50166

Name: P2MIN
Published: 2020-05-30

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR50166

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Nicholas Rego and David Koes
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Bioinformatics (2015) 31 (8): 1322-1324 doi:10.1093/bioinformatics/btu829

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Title:

P2MIN

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:

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).
PubMed: 32126231

Assembly members:

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

Natural source:

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

Experimental source:

Experimental source: Production method: recombinant technology Host organism: Escherichia coli Vector: pET-21a(+)

Entity Sequences (FASTA):

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

Data sets:

assigned_chemical_shifts
- assigned_chemical_shifts_1_P2MIN_Assigned_Chemical_Shifts
spectral_peak_list
- spectral_peak_list_1_15N_NOESY_HSQC_120ms
- spectral_peak_list_2_13C_NOESY_HSQC
theoretical_heteronucl_NOEs
- theoretical_heteronucl_NOEs_1_600NOE
- theoretical_heteronucl_NOEs_2_700NOE
theoretical_heteronucl_T1_relaxation
- theoretical_heteronucl_T1_relaxation_1_600R1
- theoretical_heteronucl_T1_relaxation_3_700R1
theoretical_heteronucl_T2_relaxation
- theoretical_heteronucl_T2_relaxation_1_600R2

Data type	Count
13C chemical shifts	502
15N chemical shifts	137
1H chemical shifts	751

Additional metadata:

Assembly
Samples and Experiments
Software
Spectrometers
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Assembly:

Entity Assembly ID	Entity Name	Entity ID
1	P2MIN	1

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

MET

THR

SER

GLU

TRP

GLU

LEU

PHE

ILE

TYR

2

LYS

LEU

LYS

ALA

ASP

LYS

TRP

CYS

ILE

GLY

3

GLU

PRO

LEU

ARG

ASN

LEU

PHE

MET

PRO

GLU

4

GLY

THR

ARG

ILE

ALA

VAL

PHE

ARG

ASP

5

ASN

GLN

LEU

HIS

PRO

SER

GLY

SER

THR

6

GLU

LEU

CYS

GLU

GLY

ASP

THR

LEU

CYS

VAL

7

MET

ALA

GLN

GLU

ARG

ASP

LEU

GLU

SER

LEU

8

SER

ARG

LEU

PHE

SER

GLU

ALA

PRO

GLU

LYS

9

ALA

SER

LEU

ALA

ARG

PHE

GLY

ASP

PHE

10

PHE

LEU

ASP

ILE

GLU

ALA

LYS

LEU

GLN

ASP

11

VAL

ALA

LEU

TYR

GLY

LEU

ASP

LEU

GLY

12

GLU

LEU

GLU

ALA

ASP

ALA

LYS

LEU

LYS

ASP

13

LEU

VAL

LEU

GLU

HIS

LEU

GLY

GLU

THR

PRO

14

VAL

LEU

GLY

ASP

TYR

PHE

GLU

TRP

HIS

GLY

15

LEU

GLN

TRP

VAL

ALA

ASP

VAL

ASP

16

TRP

LYS

VAL

THR

LYS

ILE

GLY

LEU

ARG

GLU

17

ASN

LEU

TYR

PHE

GLN

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:

Name	Sample	Sample state	Sample conditions
2D 1H-15N HSQC	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D HNCO	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D HN(CO)CA	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D HNCA	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D CBCA(CO)NH	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D HNCACB	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D HNHA	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D HNHB	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
3D HCCH-TOCSY	sample_1_P2MIN	isotropic	sample_conditions_4_P2MIN_15N_and_13C_labelled
3D CC(CO)NH	sample_1_P2MIN	isotropic	sample_conditions_4_P2MIN_15N_and_13C_labelled
3D 1H-15N NOESY	sample_1_P2MIN	isotropic	sample_conditions_1_P2MIN_15N_labelled
2D 1H-13C HSQC	sample_1_P2MIN	isotropic	sample_conditions_3_P2MIN_13C_labelled
3D 1H-13C NOESY	sample_1_P2MIN	isotropic	sample_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