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 15485

Title: Segmental isotope labeling of Npl3   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:
Npl3_RRM1, polymer, 108 residues, 12239 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):
Npl3_RRM1: MGSSHHHHHHSSGLVPRGSH MHHRQEGELSNTRLFVRPFP LDVQESELNEIFGPFGPMKE VKILNGFAFVEFEEAESAAK AIEEVHGKSFANQPLEVVYS KLPAKRYR

Data sets:
Data typeCount
13C chemical shifts237
15N chemical shifts73
1H chemical shifts513

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1RRM 11

Entities:

Entity 1, RRM 1 108 residues - 12239 Da.

Residues 1-20 represent a N-terminal sequence originated from pET28a+ vector which includes His tag

1   METGLYSERSERHISHISHISHISHISHIS
2   SERSERGLYLEUVALPROARGGLYSERHIS
3   METHISHISARGGLNGLUGLYGLULEUSER
4   ASNTHRARGLEUPHEVALARGPROPHEPRO
5   LEUASPVALGLNGLUSERGLULEUASNGLU
6   ILEPHEGLYPROPHEGLYPROMETLYSGLU
7   VALLYSILELEUASNGLYPHEALAPHEVAL
8   GLUPHEGLUGLUALAGLUSERALAALALYS
9   ALAILEGLUGLUVALHISGLYLYSSERPHE
10   ALAASNGLNPROLEUGLUVALVALTYRSER
11   LYSLEUPROALALYSARGTYRARG

Samples:

sample_1: Npl3 RRM1, [U-13C; U-15N], 1 mM; H2O 90%; D2O 10%

sample_2: Npl3 RRM1, [U-13C; U-15N], 1 mM; D2O 100%

sample_3: Npl3 RRM1, [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
3D HNCAsample_1isotropicsample_conditions_1
3D HN(CO)CAsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D CBCA(CO)NHsample_1isotropicsample_conditions_1
3D HCCH-TOCSYsample_1isotropicsample_conditions_1
3D 1H-13C NOESYsample_1isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
2D 1H-1H TOCSYsample_2isotropicsample_conditions_1
2D 1H-1H NOESYsample_2isotropicsample_conditions_1
2D 1H-13C HSQCsample_1isotropicsample_conditions_1

Software:

xwinnmr, Bruker Biospin - collection, processing

SPARKY, Goddard - chemical shift assignment, data analysis

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

NMR spectrometers:

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

Related Database Links:

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