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BMRB Entry 16481 BMRB - Biological Magnetic Resonance Bank

BMRB Entry 16481

Chem Shift validation: AVS_anomalous, AVS_full, LACS, SPARTA
BMRB Entry DOI: doi:10.13018/BMR16481

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Title:
1H, 13C and 15N resonance assignments of the hyperthermostable 264-residue endo-<beta>-1,3-glucanase LamA at 62 C
Deposition date:
2009-09-07
Original release date:
2009-11-20
Authors:
Ippel, Johannes; Koutsopoulos, Sotirios; Nabuurs, Sanne; van Berkel, Willem; van der Oost, John; van Mierlo, Carlo
Citation:

Citation: Ippel, Johannes; Koutsopoulos, Sotirios; Nabuurs, Sanne; van Berkel, Willem; van der Oost, John; van Mierlo, Carlo. "NMR characterization of a 264-residue hyperthermostable endo-beta-1,3-glucanase."  Biochem. Biophys. Res. Commun. 391, 370-375 (2010).
PubMed: 19913513

Assembly members:

Assembly members:
pfLamA, polymer, 264 residues, 30085 Da.
CA, non-polymer, 40.078 Da.

Natural source:

Natural source:   Common Name: Pyrococcus furiosus   Taxonomy ID: 2261   Superkingdom: Archaea   Kingdom: not available   Genus/species: Pyrococcus furiosus

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: DE3

Entity Sequences (FASTA):

Entity Sequences (FASTA):
pfLamA: MVPEVIEIDGKQWRLIWHDE FEGSEVNKEYWTFEKGNGIA YGIPGWGNGELEYYTENNTY IVNGTLVIEARKEIITDPNE GTFLYTSSRLKTEGKVEFSP PVVVEARIKLPKGKGLWPAF WMLGSNIREVGWPNCGEIDI MEFLGHEPRTIHGTVHGPGY SGSKGITRAYTLPEGVPDFT EDFHVFGIVWYPDKIKWYVD GTFYHEVTKEQVEAMGYEWV FDKPFYIILNLAVGGYWPGN PDATTPFPAKMVVDYVRVYS FVSG

Data typeCount
13C chemical shifts1799
15N chemical shifts526
1H chemical shifts1961

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1enzym1
2$CAA

Entities:

Entity 1, enzym 264 residues - 30085 Da.

Sequence numbers corresponds to the native pfLamA sequence without signal peptide.

1   METVALPROGLUVALILEGLUILEASPGLY
2   LYSGLNTRPARGLEUILETRPHISASPGLU
3   PHEGLUGLYSERGLUVALASNLYSGLUTYR
4   TRPTHRPHEGLULYSGLYASNGLYILEALA
5   TYRGLYILEPROGLYTRPGLYASNGLYGLU
6   LEUGLUTYRTYRTHRGLUASNASNTHRTYR
7   ILEVALASNGLYTHRLEUVALILEGLUALA
8   ARGLYSGLUILEILETHRASPPROASNGLU
9   GLYTHRPHELEUTYRTHRSERSERARGLEU
10   LYSTHRGLUGLYLYSVALGLUPHESERPRO
11   PROVALVALVALGLUALAARGILELYSLEU
12   PROLYSGLYLYSGLYLEUTRPPROALAPHE
13   TRPMETLEUGLYSERASNILEARGGLUVAL
14   GLYTRPPROASNCYSGLYGLUILEASPILE
15   METGLUPHELEUGLYHISGLUPROARGTHR
16   ILEHISGLYTHRVALHISGLYPROGLYTYR
17   SERGLYSERLYSGLYILETHRARGALATYR
18   THRLEUPROGLUGLYVALPROASPPHETHR
19   GLUASPPHEHISVALPHEGLYILEVALTRP
20   TYRPROASPLYSILELYSTRPTYRVALASP
21   GLYTHRPHETYRHISGLUVALTHRLYSGLU
22   GLNVALGLUALAMETGLYTYRGLUTRPVAL
23   PHEASPLYSPROPHETYRILEILELEUASN
24   LEUALAVALGLYGLYTYRTRPPROGLYASN
25   PROASPALATHRTHRPROPHEPROALALYS
26   METVALVALASPTYRVALARGVALTYRSER
27   PHEVALSERGLY

Entity A, $CA

Samples:

pfLamA_(50%_Ca2+)_15N_13C-labelled: pfLamA, [U-98% 13C; U-98% 15N], 1.5 mM; calcium chloride 0.75 mM; sodium phosphate 10 mM

pfLamA_(100%_Ca2+)_15N_13C-labelled: pfLamA, [U-98% 13C; U-98% 15N], 1.5 mM; sodium phosphate 10 mM; calcium chloride 10 mM

pfLamA_(50%_Ca2+)_non-labelled: pfLamA 1 mM; calcium chloride 0.5 mM; sodium phosphate 10 mM

pfLamA_(50%_Ca2+)_15N-labelled: pfLamA, [U-98% 15N], 1.5 mM; calcium chloride 0.75 mM; sodium phosphate 10 mM

sample_conditions_1: pH: 7.0; pressure: 1.0 atm; temperature: 335 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCpfLamA_(50%_Ca2+)_15N-labelledisotropicsample_conditions_1
2D 1H-13C HSQCpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
2D 1H-1H NOESYpfLamA_(50%_Ca2+)_non-labelledisotropicsample_conditions_1
2D 1H-1H TOCSYpfLamA_(50%_Ca2+)_non-labelledisotropicsample_conditions_1
3D CBCA(CO)NHpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HNCOpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HNCACBpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HBHA(CO)NHpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HCCH-TOCSYpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HN(CA)COpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D 1H-15N NOESYpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D 1H-13C NOESYpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D CNH NOESYpfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
2D 1H-15N HSQCpfLamA_(100%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D CBCA(CO)NHpfLamA_(100%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HNCOpfLamA_(100%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HNCACBpfLamA_(100%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HN(CA)COpfLamA_(100%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
2D 1H-15N HSQCpfLamA_(100%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HN(CO)CApfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1
3D HNCApfLamA_(50%_Ca2+)_15N_13C-labelledisotropicsample_conditions_1

Software:

xwinnmr, Bruker Biospin - collection, processing

NEASY/CARA, Keller and Wuthrich - chemical shift assignment, peak picking

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

SPARKY, Goddard - chemical shift assignment, peak picking

PACES, (PACES) Coggins and Zhou - chemical shift assignment

NMR spectrometers:

  • Bruker AMX 500 MHz
  • Bruker DRX 700 MHz
  • Bruker DRX 900 MHz

Related Database Links:

GB AAC25554.2 AAC25554 AAL80200 AFN04498
PDB
REF WP_011011188

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