BMRB Entry 17610

Title:
Solution structure of GppNHp-bound H-RasT35S mutant protein.
Deposition date:
2011-04-28
Original release date:
2012-05-09
Authors:
Araki, Mitsugu
Citation:

Citation: Araki, Mitsugu; Shima, Fumi; Yoshikawa, Yoko; Muraoka, Shin; Ijiri, Yuichi; Nagahara, Yuka; Shirono, Tomoya; Kataoka, Tohru; Tamura, Atsuo. "Solution structure of the state 1 conformer of GTP-bound H-Ras protein and distinct dynamic properties between the state 1 and state 2 conformers."  J. Biol. Chem. 286, 39644-39653 (2011).
PubMed: 21930707

Assembly members:

Assembly members:
GppNHp-bound H-RasT35S mutant protein, polymer, 172 residues, 18861.303 Da.
PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER, non-polymer, 522.196 Da.
MG, non-polymer, 24.305 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: pGEX-6P

Data sets:
Data typeCount
13C chemical shifts719
15N chemical shifts184
1H chemical shifts1177
T1 relaxation values130
T2 relaxation values130
heteronuclear NOE values130

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1GppNHp-bound H-RasT35S mutant protein1
2PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER2
3magnesium ions3

Entities:

Entity 1, GppNHp-bound H-RasT35S mutant protein 172 residues - 18861.303 Da.

1   GLYPROLEUGLYSERASPMETTHRGLUTYR
2   LYSLEUVALVALVALGLYALAGLYGLYVAL
3   GLYLYSSERALALEUTHRILEGLNLEUILE
4   GLNASNHISPHEVALASPGLUTYRASPPRO
5   SERILEGLUASPSERTYRARGLYSGLNVAL
6   VALILEASPGLYGLUTHRCYSLEULEUASP
7   ILELEUASPTHRALAGLYGLNGLUGLUTYR
8   SERALAMETARGASPGLNTYRMETARGTHR
9   GLYGLUGLYPHELEUCYSVALPHEALAILE
10   ASNASNTHRLYSSERPHEGLUASPILEHIS
11   GLNTYRARGGLUGLNILELYSARGVALLYS
12   ASPSERASPASPVALPROMETVALLEUVAL
13   GLYASNLYSCYSASPLEUALAALAARGTHR
14   VALGLUSERARGGLNALAGLNASPLEUALA
15   ARGSERTYRGLYILEPROTYRILEGLUTHR
16   SERALALYSTHRARGGLNGLYVALGLUASP
17   ALAPHETYRTHRLEUVALARGGLUILEARG
18   GLNHIS

Entity 2, PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER - C10 H17 N6 O13 P3 - 522.196 Da.

1   GNP

Entity 3, magnesium ions - Mg - 24.305 Da.

1   MG

Samples:

sample_1: GppNHp-bound H-RasT35S mutant protein, [U-98% 13C; U-98% 15N], 1 – 2 mM; PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER1 – 2 mM; magnesium ions 10 mM; sodium chloride 150 mM; sodium phosphate 25 mM; H20 90%; D2O 10%

sample_2: GppNHp-bound H-RasT35S mutant protein, [U-98% 13C; U-98% 15N], 1 – 2 mM; PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER1 – 2 mM; magnesium ions 10 mM; sodium chloride 150 mM; sodium phosphate 25 mM; H20 90%; D2O 10%

sample_conditions_1: ionic strength: 0.24 M; pH: 6.8; pressure: 1 atm; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-13C HSQCsample_1isotropicsample_conditions_1
3D CBCA(CO)NHsample_1isotropicsample_conditions_1
3D HNCOsample_1isotropicsample_conditions_1
3D HNCAsample_1isotropicsample_conditions_1
3D HBHA(CO)NHsample_1isotropicsample_conditions_1
3D HN(CO)CAsample_1isotropicsample_conditions_1
3D H(CCO)NHsample_1isotropicsample_conditions_1
3D HCCH-TOCSYsample_1isotropicsample_conditions_1
3D 1H-13C NOESYsample_1isotropicsample_conditions_1
3D HCCH-COSYsample_1isotropicsample_conditions_1
2D 1H-15N HSQCsample_2isotropicsample_conditions_1
3D 1H-15N NOESYsample_2isotropicsample_conditions_1
3D 1H-15N TOCSYsample_2isotropicsample_conditions_1

Software:

CYANA v2.1, Guntert, Mumenthaler and Wuthrich - structure solution

CNS v1.2, Brunger, Adams, Clore, Gros, Nilges and Read - refinement

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

SPARKY, Goddard - peak picking

NMR spectrometers:

  • Bruker DMX 750 MHz

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