BMRB

Biological Magnetic Resonance Data Bank


A Repository for Data from NMR Spectroscopy on Proteins, Peptides, Nucleic Acids, and other Biomolecules
Member of WWPDB

BMRB Entry 15487

Title: Segmental Isotope Labeling of Npl3p   PubMed: 17936301

Authors: Skrisovska, Lenka; Allain, Frederic

Citation: Skrisovska, Lenka; Allain, Frederic. "Improved segmental isotope labeling methods for the NMR study of multidomain or large proteins: application to the RRMs of Npl3p and hnRNP L"  J. Mol. Biol. 375, 151-164 (2008).

Assembly members:
Npl3p_RRM2, polymer, 111 residues, 12545 Da.

Natural source:   Common Name: baker   Taxonomy ID: 4932   Superkingdom: not available   Kingdom: not available   Genus/species: Eukaryota Fungi

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

Entity Sequences (FASTA):
Npl3p_RRM2: MGSSHHHHHHSSGLVPRGSH MSKLPAKRYRITMKNLPEGC SWQDLKDLARENSLETTFSS VNTRDFDGTGALEFPSEEIL VEALERLNNIEFRGSVITVE RDDNPPPIRRS

Data sets:
Data typeCount
13C chemical shifts230
15N chemical shifts82
1H chemical shifts501

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1Npl3p RRM21

Entities:

Entity 1, Npl3p RRM2 111 residues - 12545 Da.

1   METGLYSERSERHISHISHISHISHISHIS
2   SERSERGLYLEUVALPROARGGLYSERHIS
3   METSERLYSLEUPROALALYSARGTYRARG
4   ILETHRMETLYSASNLEUPROGLUGLYCYS
5   SERTRPGLNASPLEULYSASPLEUALAARG
6   GLUASNSERLEUGLUTHRTHRPHESERSER
7   VALASNTHRARGASPPHEASPGLYTHRGLY
8   ALALEUGLUPHEPROSERGLUGLUILELEU
9   VALGLUALALEUGLUARGLEUASNASNILE
10   GLUPHEARGGLYSERVALILETHRVALGLU
11   ARGASPASPASNPROPROPROILEARGARG
12   SER

Samples:

sample_1: Npl3p RRM2, [U-13C; U-15N], 1 mM; H2O 90%; D2O 10%

sample_2: Npl3p RRM2, [U-13C; U-15N], 1 mM; D2O 10%

sample_3: Npl3p RRM2, [U-15N], 1 mM; H2O 90%; D2O 10%

sample_conditions_1: ionic strength: 0.2 M; pH: 7; pressure: 1 atm; temperature: 303 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
2D 1H-13C HSQCsample_1isotropicsample_conditions_1
3D HNCAsample_1isotropicsample_conditions_1
3D HN(CO)CAsample_1isotropicsample_conditions_1
3D CBCA(CO)NHsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D 1H-13C NOESYsample_1isotropicsample_conditions_1
3D HCCH-TOCSYsample_1isotropicsample_conditions_1
2D 1H-1H TOCSYsample_2isotropicsample_conditions_1
2D 1H-1H NOESYsample_2isotropicsample_conditions_1

Software:

xwinnmr, Bruker Biospin - collection, processing

SPARKY, Goddard - chemical shift assignment

ATHNOS-CANDID, T. Herrmann 2005 - automated NOE assignment, automated peak picking, structure calculation

AMBER v7.0, Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Koll - refinement

NMR spectrometers:

  • Bruker DRX 500 MHz
  • Bruker DRX 600 MHz
  • Bruker Avance 900 MHz

Related Database Links:

PDB
DBJ GAA22648
EMBL CAA46817 CAA50291 CAY78932
GB AAA34818 AAB64865 AHY75384 AJP38110 AJU58232
REF NP_010720
SP Q01560
TPG DAA12270