BMRB Entry 51500

Name: Miro2 N-terminal GTPase domain bound to GTP
Published: 2022-06-27

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR51500

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Nicholas Rego and David Koes
3Dmol.js: molecular visualization with WebGL
Bioinformatics (2015) 31 (8): 1322-1324 doi:10.1093/bioinformatics/btu829

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Title:

Miro2 N-terminal GTPase domain bound to GTP

Deposition date:

2022-06-21

Original release date:

2022-06-27

Authors:

Smith, Cassandra; Jones, David

Citation:

Citation: Smith, Cassandra; Jones, David. "Backbone Chemical Shift Assignment of the N-terminal GTPase domain of Miro2 bound to GTP" Biomol. NMR Assignments ., .-..

Assembly members:

Assembly members:
entity_1, polymer, 200 residues, 22218.23 Da.
entity_GTP, non-polymer, 523.180 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: pET28a

Entity Sequences (FASTA):

Entity Sequences (FASTA):
entity_1: MGSSHHHHHHSSGLVPRGSH MRRDVRILLLGEAQVGKTSL ILSLVGEEFPEEVPPRAEEI TIPADVTPEKVPTHIVDYSE AEQTDEELREEIHKANVVCV VYDVSEEATIEKIRTKWIPL VNGGTTQGPRVPIILVGNKS DLRSGSSMEAVLPIMSQFPE IETCVECSAKNLRNISELFY YAQKAVLHPTAPLYDPEAKQ

Data sets:

assigned_chemical_shifts
- assigned_chemical_shifts_1

Data type	Count
13C chemical shifts	643
15N chemical shifts	169
1H chemical shifts	169

Additional metadata:

Assembly
Samples and Experiments
Software
Spectrometers
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Assembly:

Entity Assembly ID	Entity Name	Entity ID
1	Protein	1
2	Nucleotide	2

Entities:

Entity 1, Protein 200 residues - 22218.23 Da.

Residues 1-20 represent the non-native affinity tag for purification. The remainder represent the first residues 1-180 of the native protein.

1	MET	GLY	SER	SER	HIS	HIS	HIS	HIS	HIS	HIS
2	SER	SER	GLY	LEU	VAL	PRO	ARG	GLY	SER	HIS
3	MET	ARG	ARG	ASP	VAL	ARG	ILE	LEU	LEU	LEU
4	GLY	GLU	ALA	GLN	VAL	GLY	LYS	THR	SER	LEU
5	ILE	LEU	SER	LEU	VAL	GLY	GLU	GLU	PHE	PRO
6	GLU	GLU	VAL	PRO	PRO	ARG	ALA	GLU	GLU	ILE
7	THR	ILE	PRO	ALA	ASP	VAL	THR	PRO	GLU	LYS
8	VAL	PRO	THR	HIS	ILE	VAL	ASP	TYR	SER	GLU
9	ALA	GLU	GLN	THR	ASP	GLU	GLU	LEU	ARG	GLU
10	GLU	ILE	HIS	LYS	ALA	ASN	VAL	VAL	CYS	VAL
11	VAL	TYR	ASP	VAL	SER	GLU	GLU	ALA	THR	ILE
12	GLU	LYS	ILE	ARG	THR	LYS	TRP	ILE	PRO	LEU
13	VAL	ASN	GLY	GLY	THR	THR	GLN	GLY	PRO	ARG
14	VAL	PRO	ILE	ILE	LEU	VAL	GLY	ASN	LYS	SER
15	ASP	LEU	ARG	SER	GLY	SER	SER	MET	GLU	ALA
16	VAL	LEU	PRO	ILE	MET	SER	GLN	PHE	PRO	GLU
17	ILE	GLU	THR	CYS	VAL	GLU	CYS	SER	ALA	LYS
18	ASN	LEU	ARG	ASN	ILE	SER	GLU	LEU	PHE	TYR
19	TYR	ALA	GLN	LYS	ALA	VAL	LEU	HIS	PRO	THR
20	ALA	PRO	LEU	TYR	ASP	PRO	GLU	ALA	LYS	GLN

Entity 2, Nucleotide - C10 H16 N5 O14 P3 - 523.180 Da.

1

GTP

Samples:

sample_1: N-terminal GTPase Domain, [U-99% 13C; U-99% 15N], 350 ± 7 uM; GUANOSINE-5'-TRIPHOSPHATE 200 ± 4 uM; D2O 10 ± 0.01 %; DSS 100 ± 5 uM; sodium chloride 150 ± 1 mM; HEPES 25 ± 0.5 mM; magnesium chloride 1 ± 0.01 mM; H2O 90 ± 1 %

sample_2: N-terminal GTPase Domain, [U-99% 13C; U-99% 15N; U-80% 2H], 350 ± 7 uM; GUANOSINE-5'-TRIPHOSPHATE 200 ± 4 uM; D2O 10 ± 0.01 %; DSS 100 ± 5 uM; sodium chloride 150 ± 1 mM; HEPES 25 ± 0.5 mM; magnesium chloride 1 ± 0.01 mM; H2O 90 ± 1 %

sample_conditions_1: ionic strength: 150 mM; pH: 7.5; pressure: 1 atm; temperature: 298 K

Experiments:

Name	Sample	Sample state	Sample conditions
2D 1H-15N HSQC	sample_1	isotropic	sample_conditions_1
2D 1H-15N HSQC	sample_1	isotropic	sample_conditions_1
3D HNCACB	sample_1	isotropic	sample_conditions_1
3D CBCA(CO)NH	sample_1	isotropic	sample_conditions_1
3D C(CO)NH	sample_1	isotropic	sample_conditions_1
3D HNCO	sample_1	isotropic	sample_conditions_1
3D HN(CA)CO	sample_1	isotropic	sample_conditions_1
3D HN(CO)CA	sample_2	isotropic	sample_conditions_1
3D HN(COCA)CB	sample_2	isotropic	sample_conditions_1
3D HNCACB	sample_2	isotropic	sample_conditions_1
1D 1H	sample_1	isotropic	sample_conditions_1

Software:

VNMRj v4.2A - collection

TOPSPIN v4.1 - collection

ANALYSIS v2.5 - chemical shift assignment, data analysis, peak picking

istHMS - collection, processing

NMRPipe v10.9 - processing

NMR spectrometers:

Varian DD2 900 MHz
Bruker AVANCE NEO 600 MHz

Uniprot	Q8IXI1
AlphaFold	Q9H7M8

1	MET	GLY	SER	SER	HIS	HIS	HIS	HIS	HIS	HIS
2	SER	SER	GLY	LEU	VAL	PRO	ARG	GLY	SER	HIS
3	MET	ARG	ARG	ASP	VAL	ARG	ILE	LEU	LEU	LEU
4	GLY	GLU	ALA	GLN	VAL	GLY	LYS	THR	SER	LEU
5	ILE	LEU	SER	LEU	VAL	GLY	GLU	GLU	PHE	PRO
6	GLU	GLU	VAL	PRO	PRO	ARG	ALA	GLU	GLU	ILE
7	THR	ILE	PRO	ALA	ASP	VAL	THR	PRO	GLU	LYS
8	VAL	PRO	THR	HIS	ILE	VAL	ASP	TYR	SER	GLU
9	ALA	GLU	GLN	THR	ASP	GLU	GLU	LEU	ARG	GLU
10	GLU	ILE	HIS	LYS	ALA	ASN	VAL	VAL	CYS	VAL
11	VAL	TYR	ASP	VAL	SER	GLU	GLU	ALA	THR	ILE
12	GLU	LYS	ILE	ARG	THR	LYS	TRP	ILE	PRO	LEU
13	VAL	ASN	GLY	GLY	THR	THR	GLN	GLY	PRO	ARG
14	VAL	PRO	ILE	ILE	LEU	VAL	GLY	ASN	LYS	SER
15	ASP	LEU	ARG	SER	GLY	SER	SER	MET	GLU	ALA
16	VAL	LEU	PRO	ILE	MET	SER	GLN	PHE	PRO	GLU
17	ILE	GLU	THR	CYS	VAL	GLU	CYS	SER	ALA	LYS
18	ASN	LEU	ARG	ASN	ILE	SER	GLU	LEU	PHE	TYR
19	TYR	ALA	GLN	LYS	ALA	VAL	LEU	HIS	PRO	THR
20	ALA	PRO	LEU	TYR	ASP	PRO	GLU	ALA	LYS	GLN

1	MET	GLY	SER	SER	HIS	HIS	HIS	HIS	HIS	HIS
2	SER	SER	GLY	LEU	VAL	PRO	ARG	GLY	SER	HIS
3	MET	ARG	ARG	ASP	VAL	ARG	ILE	LEU	LEU	LEU
4	GLY	GLU	ALA	GLN	VAL	GLY	LYS	THR	SER	LEU
5	ILE	LEU	SER	LEU	VAL	GLY	GLU	GLU	PHE	PRO
6	GLU	GLU	VAL	PRO	PRO	ARG	ALA	GLU	GLU	ILE
7	THR	ILE	PRO	ALA	ASP	VAL	THR	PRO	GLU	LYS
8	VAL	PRO	THR	HIS	ILE	VAL	ASP	TYR	SER	GLU
9	ALA	GLU	GLN	THR	ASP	GLU	GLU	LEU	ARG	GLU
10	GLU	ILE	HIS	LYS	ALA	ASN	VAL	VAL	CYS	VAL
11	VAL	TYR	ASP	VAL	SER	GLU	GLU	ALA	THR	ILE
12	GLU	LYS	ILE	ARG	THR	LYS	TRP	ILE	PRO	LEU
13	VAL	ASN	GLY	GLY	THR	THR	GLN	GLY	PRO	ARG
14	VAL	PRO	ILE	ILE	LEU	VAL	GLY	ASN	LYS	SER
15	ASP	LEU	ARG	SER	GLY	SER	SER	MET	GLU	ALA
16	VAL	LEU	PRO	ILE	MET	SER	GLN	PHE	PRO	GLU
17	ILE	GLU	THR	CYS	VAL	GLU	CYS	SER	ALA	LYS
18	ASN	LEU	ARG	ASN	ILE	SER	GLU	LEU	PHE	TYR
19	TYR	ALA	GLN	LYS	ALA	VAL	LEU	HIS	PRO	THR
20	ALA	PRO	LEU	TYR	ASP	PRO	GLU	ALA	LYS	GLN