BMRB Entry 15442

Title:
Multienzyme Docking in Hybrid Megasynthetases
Deposition date:
2007-08-24
Original release date:
2008-06-26
Authors:
Richter, Carsten; Nietlispach, Daniel; Broadhurst, Richard; Weissman, Kira
Citation:

Citation: Richter, Carsten; Nietlispach, Daniel; Broadhurst, Richard; Weissman, Kira. "Multienzyme docking in hybrid megasynthetases"  Nat. Chem. Biol. 4, 75-81 (2008).
PubMed: 18066054

Assembly members:

Assembly members:
TubCdd, polymer, 78 residues, 8132.495 Da.

Natural source:

Natural source:   Common Name: Angiococcus disciformis   Taxonomy ID: 38   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Angiococcus disciformis

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pGEX-6P-1

Entity Sequences (FASTA):

Data sets:
Data typeCount
13C chemical shifts308
15N chemical shifts75
1H chemical shifts515

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1TubCdd A1
2TubCdd B1

Entities:

Entity 1, TubCdd A 78 residues - 8132.495 Da.

The first four residues represent the remnant of the Precission protease cleavage sequence that separated the polypeptide from its N-terminal glutathoine-S-transferase fusion partner.

1   GLYPROLEUGLYSERSERALAGLYALALEU
2   LEUALAHISALAALASERLEUGLYVALARG
3   LEUTRPVALGLUGLYGLUARGLEUARGPHE
4   GLNALAPROPROGLYVALMETTHRPROGLU
5   LEUGLNSERARGLEUGLYGLYALAARGHIS
6   GLULEUILEALALEULEUARGGLNLEUGLN
7   PROSERSERGLNGLYGLYSERLEULEUALA
8   PROVALALAARGASNGLYARGLEU

Samples:

sample_1: TubCdd, [U-100% 13C; U-100% 15N], 1 mM; TSP 20 uM; sodium azide 0.1%; sodium phosphate 50 mM; sodium chloride 100 mM; H2O 90%; D2O 10%

sample_2: TubCdd, [U-100% 13C; U-100% 15N], 0.5 mM; TubCdd 0.5 mM; TSP 20 uM; sodium azide 0.1%; sodium phosphate 50 mM; sodium chloride 100 mM; TubCdd 0.5 mM; H2O 90%; D2O 10%

sample_conditions_1: ionic strength: 0.15 M; pH: 6; pressure: 1 atm; temperature: 308 K

Experiments:

NameSampleSample stateSample conditions
3D 1H-13C NOESYsample_1isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D 13C-separated 12C-selected NOESYsample_2isotropicsample_conditions_1
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
2D 1H-13C HSQCsample_1isotropicsample_conditions_1
3D CBCA(CO)NHsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D HCCH-TOCSYsample_1isotropicsample_conditions_1
3D HBHA(CO)NHsample_1isotropicsample_conditions_1
3D HNCOsample_1isotropicsample_conditions_1
3D 1H-15N TOCSYsample_1isotropicsample_conditions_1

Software:

ARIA v1.2, Linge, O'Donoghue and Nilges - structure solution

AZARA, Boucher - processing

ANALYSIS v1.0.15, Vranken, Boucher, Stevens, Fogh, Pajon, Llinas, Ulrich, Markley, Ionides and Laue - chemical shift assignment, data analysis, peak picking

NMR spectrometers:

  • Bruker DRX 500 MHz
  • Bruker DRX 800 MHz

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