BMRB Entry 19725

Name: Backbone 1H and 15N Chemical Shift Assignments of a Double Y to E Mutant of the Cytoplasmic Tail of Ig-alpha (CD79a) in a Heterodimeric Construct
Published: 2014-02-06

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR19725

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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:

Backbone 1H and 15N Chemical Shift Assignments of a Double Y to E Mutant of the Cytoplasmic Tail of Ig-alpha (CD79a) in a Heterodimeric Construct

Deposition date:

2014-01-08

Original release date:

2014-02-06

Authors:

Bentrop, Detlef; Salavei, Pavel; Reth, Michael; Fakler, Bernd

Citation:

Citation: Salavei, Pavel; Bentrop, Detlef; Reth, Michael; Fakler, Bernd. "A heterodimeric construct mimicking the cytoplasmic region of the B-cell antigen receptor complex-associated protein" .

Assembly members:

Assembly members:
cytoplasmic_Ig-alpha_polypeptide_Y_to_E_double_mutant, polymer, 99 residues, Formula weight is not available
cytoplasmic_Ig-beta_polypeptide, polymer, 86 residues, Formula weight is not available

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: pET23a

Entity Sequences (FASTA):

Entity Sequences (FASTA):
cytoplasmic_Ig-alpha_polypeptide_Y_to_E_double_mutant: SGGVAQLRERVKTLRAQNYE LESEVQRLREQVAQLSGGRK RWQNEKLGLDAGDEYEDENL EEGLNLDDCSMEEDISRGLQ GTYQDVGSLNIGDVQLEKP
cytoplasmic_Ig-beta_polypeptide: SGGVDELQAEVDQLQDENYA LKTKVAQLRKKVEKLSGGDK DDSKAGMEEDHTYEGLDIDQ TATYEDIVTLRTGEVKWSVG EHPGQE

Data sets:

assigned_chemical_shifts
- assigned_chem_shift_list_1
heteronucl_NOEs
- heteronuclear_noe_list_1

Data type	Count
15N chemical shifts	95
1H chemical shifts	190
heteronuclear NOE values	92

Additional metadata:

Assembly
Samples and Experiments
Software
Spectrometers
Hide all

Assembly:

Entity Assembly ID	Entity Name	Entity ID
1	Ig-alpha YE variant Ig-beta construct, subunit 1	1
2	Ig-alpha YE variant Ig-beta construct, subunit 2	2

Entities:

Entity 1, Ig-alpha YE variant Ig-beta construct, subunit 1 99 residues - Formula weight is not available

Residues 1-3 represent a linker to an affinity tag that was removed by proteolysis. Residues 4-35 are a coiled-coil heterodimerization domain (A2, see citation 3 and 1, respectively). Residues 36-38 represent a linker. Residues 39-99 are a Y to E variant of the cytoplasmic region of human Ig-alpha and contain an immunoreceptor tyrosine-based activation motif (ITAM). Both tyrosines of this motif (Y61 and Y72) were mutated to glutamate.

1

SER

GLY

VAL

ALA

GLN

LEU

ARG

GLU

ARG

2

VAL

LYS

THR

LEU

ARG

ALA

GLN

ASN

TYR

GLU

3

LEU

GLU

SER

GLU

VAL

GLN

ARG

LEU

ARG

GLU

4

GLN

VAL

ALA

GLN

LEU

SER

GLY

ARG

LYS

5

ARG

TRP

GLN

ASN

GLU

LYS

LEU

GLY

LEU

ASP

6

ALA

GLY

ASP

GLU

TYR

GLU

ASP

GLU

ASN

LEU

7

GLU

GLY

LEU

ASN

LEU

ASP

CYS

SER

8

MET

GLU

ASP

ILE

SER

ARG

GLY

LEU

GLN

9

GLY

THR

TYR

GLN

ASP

VAL

GLY

SER

LEU

ASN

10

ILE

GLY

ASP

VAL

GLN

LEU

GLU

LYS

PRO

Entity 2, Ig-alpha YE variant Ig-beta construct, subunit 2 86 residues - Formula weight is not available

Residues 1-3 represent a linker to an affinity tag that was removed by proteolysis. Residues 4-35 are a coiled-coil heterodimerization domain (B1, see citation 3 and 1, respectively). Residues 36-38 represent a linker. Residues 39-86 are the cytoplasmic region of human Ig-beta and contain an immunoreceptor tyrosine-based activation motif (ITAM).

1

SER

GLY

VAL

ASP

GLU

LEU

GLN

ALA

GLU

2

VAL

ASP

GLN

LEU

GLN

ASP

GLU

ASN

TYR

ALA

3

LEU

LYS

THR

LYS

VAL

ALA

GLN

LEU

ARG

LYS

4

LYS

VAL

GLU

LYS

LEU

SER

GLY

ASP

LYS

5

ASP

SER

LYS

ALA

GLY

MET

GLU

ASP

6

HIS

THR

TYR

GLU

GLY

LEU

ASP

ILE

ASP

GLN

7

THR

ALA

THR

TYR

GLU

ASP

ILE

VAL

THR

LEU

8

ARG

THR

GLY

GLU

VAL

LYS

TRP

SER

VAL

GLY

9

GLU

HIS

PRO

GLY

GLN

GLU

Samples:

sample_1: cytoplasmic Ig-alpha polypeptide Y to E double mutant, [U-98% 15N], 0.45 mM; cytoplasmic Ig-beta polypeptide 0.45 mM; sodium phosphate 50 mM

sample_conditions_1: pH: 6.8; pressure: 1 atm; temperature: 298 K

Experiments:

Name	Sample	Sample state	Sample conditions
2D 1H-15N HSQC	sample_1	isotropic	sample_conditions_1
3D 1H-15N NOESY	sample_1	isotropic	sample_conditions_1
3D HNHA	sample_1	isotropic	sample_conditions_1
2D 1H-15N heteroNOE	sample_1	isotropic	sample_conditions_1

Software:

TOPSPIN v2.1, Bruker Biospin - NMR data acquisition

CARA vv1.8.4.2, Keller and Wuthrich - chemical shift assignment

NMR spectrometers:

Bruker Avance 600 MHz

EMBL	S46706.1 M80461.1
BMRB	19723 19724 18884 19650 19651 19723 19724
GB	EDM06394 EGV99168 ELW67499
REF	XP_003501912 XP_003913329 XP_006041680 XP_006041681 XP_006146855