BMRB Entry 17323

Name: Solution NMR Structure of Med25(391-543) Comprising the Activator-Interacting Domain (ACID) of Human Mediator Subuniti 25. Northeast Structural Genomics Consortium Target HR6188A
Published: 2010-12-22

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PDB ID: 2l6u
Entry in NMR Restraints Grid
Validation report in NRG-CING
Chem Shift validation: AVS_full, LACS
BMRB Entry DOI: doi:10.13018/BMR17323
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Bioinformatics (2015) 31 (8): 1322-1324 doi:10.1093/bioinformatics/btu829

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Title:

Solution NMR Structure of Med25(391-543) Comprising the Activator-Interacting Domain (ACID) of Human Mediator Subuniti 25. Northeast Structural Genomics Consortium Target HR6188A

Deposition date:

2010-11-24

Original release date:

2010-12-22

Authors:

Eletsky, Alexander; Ryuechan, William; Sukumaran, Dinesh; Shastry, Ritu; Ciccosanti, Colleen; Janjua, Haleema; Acton, Thomas; Xiao, Rong; Everett, John; Montelione, Gaetano; Szyperski, Thomas

Citation:

Citation: Eletsky, Alexander; Ruyechan, William; Xiao, Rong; Acton, Thomas; Montelione, Gaetano; Szyperski, Thomas. "Solution NMR structure of MED25(391-543) comprising the activator-interacting domain (ACID) of human mediator subunit 25." J. Struct. Funct. Genomics 12, 159-166 (2011).
PubMed: 21785987

Assembly members:

Assembly members:
HR6188A, polymer, 163 residues, 18637.756 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: pET 14-15C

Entity Sequences (FASTA):

Entity Sequences (FASTA):
HR6188A: MGHHHHHHSHGQQSVSNKLL AWSGVLEWQEKPKPASVDAN TKLTRSLPCQVYVNHGENLK TEQWPQKLIMQLIPQQLLTT LGPLFRNSRMVQFHFTNKDL ESLKGLYRIMGNGFAGCVHF PHTAPCEVRVLMLLYSSKKK IFMGLIPYDQSGFVNGIRQV ITN

Data sets:

assigned_chemical_shifts
- assigned_chem_shift_list_1

Data type	Count
13C chemical shifts	721
15N chemical shifts	172
1H chemical shifts	1160

Time Domain Data

Additional metadata:

Assembly
Samples and Experiments
Software
Spectrometers
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Assembly:

Entity Assembly ID	Entity Name	Entity ID
1	HR6188A	1

Entities:

Entity 1, HR6188A 163 residues - 18637.756 Da.

residues 381-390 represent a non-native affinity tag

1

MET

GLY

HIS

SER

HIS

2

GLY

GLN

SER

VAL

SER

ASN

LYS

LEU

3

ALA

TRP

SER

GLY

VAL

LEU

GLU

TRP

GLN

GLU

4

LYS

PRO

LYS

PRO

ALA

SER

VAL

ASP

ALA

ASN

5

THR

LYS

LEU

THR

ARG

SER

LEU

PRO

CYS

GLN

6

VAL

TYR

VAL

ASN

HIS

GLY

GLU

ASN

LEU

LYS

7

THR

GLU

GLN

TRP

PRO

GLN

LYS

LEU

ILE

MET

8

GLN

LEU

ILE

PRO

GLN

LEU

THR

9

LEU

GLY

PRO

LEU

PHE

ARG

ASN

SER

ARG

MET

10

VAL

GLN

PHE

HIS

PHE

THR

ASN

LYS

ASP

LEU

11

GLU

SER

LEU

LYS

GLY

LEU

TYR

ARG

ILE

MET

12

GLY

ASN

GLY

PHE

ALA

GLY

CYS

VAL

HIS

PHE

13

PRO

HIS

THR

ALA

PRO

CYS

GLU

VAL

ARG

VAL

14

LEU

MET

LEU

TYR

SER

LYS

15

ILE

PHE

MET

GLY

LEU

ILE

PRO

TYR

ASP

GLN

16

SER

GLY

PHE

VAL

ASN

GLY

ILE

ARG

GLN

VAL

17

ILE

THR

ASN

Samples:

HR6188A_NC: HR6188A, [U-100% 13C; U-100% 15N], 0.7 mM; MES 20 mM; sodium chloride 100 mM; calcium chloride 5 mM; DTT 10 mM; sodium azide 0.02%; D2O 10%; H2O 90%

HR6188A_NC5: HR6188A, [U-5% 13C; U-100% 15N], 0.4 mM; MES 20 mM; sodium chloride 100 mM; calcium chloride 5 mM; DTT 10 mM; sodium azide 0.02%; D2O 10%; H2O 90%

sample_conditions_1: pH: 6.5; pressure: 1 atm; temperature: 298 K

Experiments:

Name	Sample	Sample state	Sample conditions
2D 1H-15N HSQC	HR6188A_NC	isotropic	sample_conditions_1
2D 1H-13C HSQC aliphatic	HR6188A_NC	isotropic	sample_conditions_1
3D HNCO	HR6188A_NC	isotropic	sample_conditions_1
3D CBCA(CO)NH	HR6188A_NC	isotropic	sample_conditions_1
3D HNCACB	HR6188A_NC	isotropic	sample_conditions_1
3D HN(CA)CO	HR6188A_NC	isotropic	sample_conditions_1
3D simutaneous 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY	HR6188A_NC	isotropic	sample_conditions_1
3D HBHA(CO)NH	HR6188A_NC	isotropic	sample_conditions_1
3D HNCA	HR6188A_NC	isotropic	sample_conditions_1
2D 1H-13C CT-HSQC aliphatic	HR6188A_NC	isotropic	sample_conditions_1
2D 1H-13C CT-HSQC aromatic	HR6188A_NC	isotropic	sample_conditions_1
1D 15N T1	HR6188A_NC	isotropic	sample_conditions_1
1D 15N T2	HR6188A_NC	isotropic	sample_conditions_1
3D (H)CCH-TOCSY aliphatic	HR6188A_NC	isotropic	sample_conditions_1
3D (H)CCH-COSY aliphatic	HR6188A_NC	isotropic	sample_conditions_1
3D (H)CCH-COSY aromatic	HR6188A_NC	isotropic	sample_conditions_1
2D 1H-15N LR-HSQC for histidines	HR6188A_NC	isotropic	sample_conditions_1
2D (HB)CB(CGCDCE)HDHE	HR6188A_NC	isotropic	sample_conditions_1
2D 1H-13C CT-HSQC methyl	HR6188A_NC5	isotropic	sample_conditions_1

Software:

CNS v1.2, Brunger, Adams, Clore, Gros, Nilges and Read - refinement,structure solution,geometry optimization

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

AutoStruct v2.2.1, Huang, Tejero, Powers and Montelione - data analysis,refinement

AutoAssign v2.3.0, Zimmerman, Moseley, Kulikowski and Montelione - data analysis,chemical shift assignment

XEASY v1.3.13, Bartels et al. - data analysis

TOPSPIN v2.1, Bruker Biospin - collection, processing

VNMRJ v2.2D, Varian - collection

PROSA v6.4, Guntert - processing

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

TALOS+ v1.2009.0721.18, Shen, Cornilescu, Delaglio and Bax - geometry optimization

NMR spectrometers:

Bruker Avance 900 MHz
Varian INOVA 750 MHz

BMRB	17139
PDB	2KY6 2L23 2L6U 2XNF
DBJ	BAE29135 BAF82149 BAG51583 BAG61008
EMBL	CAB66680 CAE84581
GB	AAG15589 AAH21333 AAH24312 AAH31138 AAH65297
REF	NP_001075914 NP_001163897 NP_083641 NP_112235 XP_001115473
SP	A2VE44 Q71SY5 Q8VCB2
TPG	DAA19611
AlphaFold	A2VE44 Q71SY5 Q8VCB2