BMRB Entry 26688

Title:
Backbone 1H, 13C, and 15N Chemical Shift Assignments of Guanylyl Cyclase Activator Protein 1 (GCAP1) mutant V77E in a Ca2+-free/Mg2+-bound Activator State
Deposition date:
2015-10-08
Original release date:
2015-12-28
Authors:
Lim, Sunghyuk; Ames, James
Citation:

Citation: Lim, Sunghyuk; Peshenko, Igor; Olshevskaya, Elena; Dizhoor, Alexander; Ames, James. "Structure of Guanylyl Cyclase Activator Protein 1 (GCAP1) Mutant V77E in a Ca2+-free/Mg2+-bound Activator State"  J. Biol. Chem. 291, 4429-4441 (2016).
PubMed: 26703466

Assembly members:

Assembly members:
GCAP1_(V77E)_mutant, polymer, 204 residues, Formula weight is not available
entity_MYR, non-polymer, 228.371 Da.
entity_MG, non-polymer, 24.305 Da.

Natural source:

Natural source:   Common Name: cow   Taxonomy ID: 9913   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Bos taurus

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pET11

Data sets:
Data typeCount
13C chemical shifts283
15N chemical shifts113
1H chemical shifts188

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1GCAP1 (V77E) mutant1
2myristic acid2
3Mg2+ ion3

Entities:

Entity 1, GCAP1 (V77E) mutant 204 residues - Formula weight is not available

A myristoyl group is attached to the N-terminal of the first residue glycine.

1   GLYASNILEMETASPGLYLYSSERVALGLU
2   GLULEUSERSERTHRGLUCYSHISGLNTRP
3   TYRLYSLYSPHEMETTHRGLUCYSPROSER
4   GLYGLNLEUTHRLEUTYRGLUPHEARGGLN
5   PHEPHEGLYLEULYSASNLEUSERPROTRP
6   ALASERGLNTYRVALGLUGLNMETPHEGLU
7   THRPHEASPPHEASNLYSASPGLYTYRILE
8   ASPPHEMETGLUTYRGLUALAALALEUSER
9   LEUVALLEULYSGLYLYSVALGLUGLNLYS
10   LEUARGTRPTYRPHELYSLEUTYRASPVAL
11   ASPGLYASNGLYCYSILEASPARGASPGLU
12   LEULEUTHRILEILEARGALAILEARGALA
13   ILEASNPROCYSSERASPSERTHRMETTHR
14   ALAGLUGLUPHETHRASPTHRVALPHESER
15   LYSILEASPVALASNGLYASPGLYGLULEU
16   SERLEUGLUGLUPHEMETGLUGLYVALGLN
17   LYSASPGLNMETLEULEUASPTHRLEUTHR
18   ARGSERLEUASPLEUTHRARGILEVALARG
19   ARGLEUGLNASNGLYGLUGLNASPGLUGLU
20   GLYALASERGLYARGGLUTHRGLUALAALA
21   GLUALAASPGLY

Entity 2, myristic acid - C14 H28 O2 - 228.371 Da.

1   MYR

Entity 3, Mg2+ ion - Mg - 24.305 Da.

1   MG

Samples:

sample_1: TRIS, [U-100% 2H], 5 mM; DTT, [U-100% 2H], 5 mM; GCAP1 (V77E) mutant, [U-100% 15N], 0.5 mM; MgCl2 5 mM; H2O 93%; D2O, [U-100% 2H], 7%

sample_2: TRIS, [U-100% 2H], 5 mM; DTT, [U-100% 2H], 5 mM; GCAP1 (V77E) mutant, [U-100% 13C; U-100% 15N], 0.5 mM; MgCl2 5 mM; H2O 93%; D2O, [U-100% 2H], 7%

sample_3: TRIS, [U-100% 2H], 5 mM; DTT, [U-100% 2H], 5 mM; GCAP1 (V77E) mutant, [U-13C; U-15N; U-2H], 0.5 mM; MgCl2 5 mM; H2O 93%; D2O, [U-100% 2H], 7%

sample_conditions_1: ionic strength: 20 mM; pH: 7.4; pressure: 1 atm; temperature: 320 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
3D HNCOsample_3isotropicsample_conditions_1
3D HNCAsample_2isotropicsample_conditions_1
3D HNCACBsample_2isotropicsample_conditions_1
3D CBCA(CO)NHsample_2isotropicsample_conditions_1
3D HBHA(CO)NHsample_2isotropicsample_conditions_1
3D 1H-15N TOCSYsample_1isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D HNCACBsample_3isotropicsample_conditions_1

Software:

NMRPipe, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - processing

SPARKY, Goddard - chemical shift assignment

NMR spectrometers:

  • Bruker Avance 800 MHz

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