BMRB Entry 19725

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):

Data sets:
Data typeCount
15N chemical shifts95
1H chemical shifts190
heteronuclear NOE values92

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1Ig-alpha YE variant Ig-beta construct, subunit 11
2Ig-alpha YE variant Ig-beta construct, subunit 22

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   SERGLYGLYVALALAGLNLEUARGGLUARG
2   VALLYSTHRLEUARGALAGLNASNTYRGLU
3   LEUGLUSERGLUVALGLNARGLEUARGGLU
4   GLNVALALAGLNLEUSERGLYGLYARGLYS
5   ARGTRPGLNASNGLULYSLEUGLYLEUASP
6   ALAGLYASPGLUTYRGLUASPGLUASNLEU
7   GLUGLUGLYLEUASNLEUASPASPCYSSER
8   METGLUGLUASPILESERARGGLYLEUGLN
9   GLYTHRTYRGLNASPVALGLYSERLEUASN
10   ILEGLYASPVALGLNLEUGLULYSPRO

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   SERGLYGLYVALASPGLULEUGLNALAGLU
2   VALASPGLNLEUGLNASPGLUASNTYRALA
3   LEULYSTHRLYSVALALAGLNLEUARGLYS
4   LYSVALGLULYSLEUSERGLYGLYASPLYS
5   ASPASPSERLYSALAGLYMETGLUGLUASP
6   HISTHRTYRGLUGLYLEUASPILEASPGLN
7   THRALATHRTYRGLUASPILEVALTHRLEU
8   ARGTHRGLYGLUVALLYSTRPSERVALGLY
9   GLUHISPROGLYGLNGLU

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:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D HNHAsample_1isotropicsample_conditions_1
2D 1H-15N heteroNOEsample_1isotropicsample_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

Related Database Links:

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

Download HSQC peak lists in one of the following formats:
CSV: Backbone or all simulated peaks
SPARKY: Backbone or all simulated peaks