BMRB

Biological Magnetic Resonance Data Bank


A Repository for Data from NMR Spectroscopy on Proteins, Peptides, Nucleic Acids, and other Biomolecules
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BMRB Entry 18044

Title: Computational design of an eight-stranded (beta/alpha)-barrel from fragments of different folds   PubMed: 22329686

Authors: Coles, Murray; Truffault, Vincent; Eisenbeis, Simone; Proffitt, William; Meiler, Jens; Hocker, Birte

Citation: Eisenbeis, Simone; Proffitt, William; Coles, Murray; Truffault, Vincent; Shanmugarantam, Sooruban; Meiler, Jens; Hocker, Birte. "Potential of fragment recombination for rational design of proteins"  J. Am. Chem. Soc. 134, 4019-4022 (2012).

Assembly members:
CheYHisF-sfr_RM, polymer, 234 residues, 25607.779 Da.

Natural source:   Common Name: not available   Taxonomy ID: not available   Superkingdom: not available   Kingdom: not available   Genus/species: not available not available

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

Entity Sequences (FASTA):
CheYHisF-sfr_RM: MGKIVLIVDDATNGREAVEK YKELKPDIVTMDITMPEMNG IDAIKEIMKIDPNAKIIVCS AMGQQAMVIEAIKAGAKGFI VNTAAVENPSLITQLAQTFG SQAVVVAIDAKRVDGEFMVF TYSGKKNTGILLRDWVVEVE KRGAGEILLTSIDRDGTKSG YDTEMIRFVRPLTTLPIIAS GGAGKMEHFLEAFLAGADAA AAASVFHFREIDGRELKEYL KKHGVNLEHHHHHH

Data sets:
Data typeCount
13C chemical shifts904
15N chemical shifts217
1H chemical shifts1578

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1CheYHisF-sfr_RM1

Entities:

Entity 1, CheYHisF-sfr_RM 234 residues - 25607.779 Da.

The protein is an engineered chimera between CheY (1-11 and 34-103 from 1TMY) and HisF (103-247 from 1THF) from Thermotoga maritima with five point mutations (R4I, D78G, I95L, L201A, V213G). The last 7 residues represent a purification tag.

1   METGLYLYSILEVALLEUILEVALASPASP
2   ALATHRASNGLYARGGLUALAVALGLULYS
3   TYRLYSGLULEULYSPROASPILEVALTHR
4   METASPILETHRMETPROGLUMETASNGLY
5   ILEASPALAILELYSGLUILEMETLYSILE
6   ASPPROASNALALYSILEILEVALCYSSER
7   ALAMETGLYGLNGLNALAMETVALILEGLU
8   ALAILELYSALAGLYALALYSGLYPHEILE
9   VALASNTHRALAALAVALGLUASNPROSER
10   LEUILETHRGLNLEUALAGLNTHRPHEGLY
11   SERGLNALAVALVALVALALAILEASPALA
12   LYSARGVALASPGLYGLUPHEMETVALPHE
13   THRTYRSERGLYLYSLYSASNTHRGLYILE
14   LEULEUARGASPTRPVALVALGLUVALGLU
15   LYSARGGLYALAGLYGLUILELEULEUTHR
16   SERILEASPARGASPGLYTHRLYSSERGLY
17   TYRASPTHRGLUMETILEARGPHEVALARG
18   PROLEUTHRTHRLEUPROILEILEALASER
19   GLYGLYALAGLYLYSMETGLUHISPHELEU
20   GLUALAPHELEUALAGLYALAASPALAALA
21   ALAALAALASERVALPHEHISPHEARGGLU
22   ILEASPGLYARGGLULEULYSGLUTYRLEU
23   LYSLYSHISGLYVALASNLEUGLUHISHIS
24   HISHISHISHIS

Samples:

15N-labelled: CheYHisF-sfr_RM, [U-100% 15N], 0.5 mM; potassium phosphate 50 mM; potassium chloride 300 mM; H2O 90%; D2O 10%

double-labelled: CheYHisF-sfr_RM, [U-100% 13C; U-100% 15N], 0.5 mM; potassium phosphate 50 mM; potassium chloride 300 mM; H2O 90%; D2O 10%

sample_conditions_1: ionic strength: 350 mM; pH: 7.5; pressure: 1 atm; temperature: 313 K

Experiments:

NameSampleSample stateSample conditions
3D HNCOdouble-labelledisotropicsample_conditions_1
3D HNCAdouble-labelledisotropicsample_conditions_1
3D HNCACBdouble-labelledisotropicsample_conditions_1
3D C(CO)NHdouble-labelledisotropicsample_conditions_1
3D CCH-TOCSYdouble-labelledisotropicsample_conditions_1
3D CCH-COSYdouble-labelledisotropicsample_conditions_1
2D PLUSH-TACSYdouble-labelledisotropicsample_conditions_1
3D HNHA15N-labelledisotropicsample_conditions_1
3D HNHB15N-labelledisotropicsample_conditions_1
3D 1H-15N NOESY15N-labelledisotropicsample_conditions_1
3D 1H-13C NOESYdouble-labelledisotropicsample_conditions_1
3D CNH-NOESYdouble-labelledisotropicsample_conditions_1
3D CCH-NOESYdouble-labelledisotropicsample_conditions_1
3D NNH-NOESY15N-labelledisotropicsample_conditions_1
2D 15N-filtered 1H-1H NOESY15N-labelledisotropicsample_conditions_1

Software:

TOPSPIN, Bruker Biospin - collection, processing

SPARKY, Goddard - chemical shift assignment, data analysis

X-PLOR NIH v2.21, Schwieters, Kuszewski, Tjandra and Clore - refinement, structure solution

NMR spectrometers:

  • Bruker AvanceIII 600 MHz
  • Bruker AvanceIII 800 MHz

Related Database Links:

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