BMRB Entry 16483

Title:
Fusion construct of CR17 from LRP-1 and ApoE residues 130-149
Deposition date:
2009-09-08
Original release date:
2010-02-02
Authors:
Guttman, Miklos; Komives, Elizabeth
Citation:

Citation: Guttman, Miklos; Prieto, J. Helena; Croy, Johnny; Komives, Elizabeth. "Decoding of lipoprotein-receptor interactions: properties of ligand binding modules governing interactions with apolipoprotein E."  Biochemistry 49, 1207-1216 (2010).
PubMed: 20030366

Assembly members:

Assembly members:
CR17, polymer, 80 residues, 8399.479 Da.
apoE, polymer, 23 residues, 8399.479 Da.
CA, non-polymer, 40.078 Da.

Natural source:

Natural source:   Common Name: Human   Taxonomy ID: 9606   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Homo sapiens

Experimental source:

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

Data sets:
Data typeCount
13C chemical shifts350
15N chemical shifts89
1H chemical shifts651
heteronuclear NOE values60
T1 relaxation values46
T2 relaxation values44

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1CR171
2ApoE2
3CALCIUM ION3

Entities:

Entity 1, CR17 80 residues - 8399.479 Da.

Residues 3-50 correspond to mature human LRP-1 (2751-2798), followed by an 8 residue linker (GSGSGSGS) and human mature ApoE residues 130-149

1   GLYSERLYSLEUGLUGLYLYSTHRCYSGLY
2   PROSERSERPHESERCYSPROGLYTHRHIS
3   VALCYSVALPROGLUARGTRPLEUCYSASP
4   GLYASPLYSASPCYSALAASPGLYALAASP
5   GLUSERILEALAALAGLYCYSLEUTYRASN
6   SERTHRGLYSERGLYSERGLYSERGLYSER
7   THRGLUGLULEUARGVALARGLEUALASER
8   HISLEUARGLYSLEUARGLYSARGLEULEU

Entity 2, ApoE 23 residues - 8399.479 Da.

N terminal GSY overhang from expression vector followed by human mature apoE residues 130-149

1   GLYSERTYRTHRGLUGLULEUARGVALARG
2   LEUALASERHISLEUARGLYSLEUARGLYS
3   ARGLEULEU

Entity 3, CALCIUM ION - Ca - 40.078 Da.

1   CA

Samples:

sample_1: HEPES, [U-99% 2H], 20 mM; sodium chloride 50 mM; sodium azide 3 mM; CALCIUM ION 5 mM; entity_1, [U-99% 13C; U-99% 15N], 0.8 mM; H2O 90%; D2O 10%

sample_2: HEPES, [U-99% 2H], 20 mM; sodium chloride 50 mM; sodium azide 3 mM; CALCIUM ION 5 mM; entity_1, [U-99% 13C; U-99% 15N], 0.8 mM; D2O 100%

sample_3: HEPES, [U-99% 2H], 20 mM; sodium chloride 50 mM; sodium azide 3 mM; CALCIUM ION 5 mM; entity_1, [U-99% 15N], 0.8 mM; H2O 90%; D2O 10%

sample_4: HEPES, [U-99% 2H], 20 mM; sodium chloride 150 mM; sodium azide 3 mM; EDTA, [U-99% 2H], 2 mM; entity_2, [U-99% 15N], 0.5 mM; H2O 90%; D2O 10%

sample_5: HEPES, [U-99% 2H], 20 mM; sodium chloride 150 mM; sodium azide 3 mM; EDTA, [U-99% 2H], 2 mM; entity_2, [U-99% 13C; U-99% 15N], 0.5 mM; H2O 90%; D2O 10%

sample_conditions_1: ionic strength: 0.05 M; pH: 7.45; pressure: 1 atm; temperature: 298 K

sample_conditions_2: ionic strength: 0.15 M; pH: 7.45; pressure: 1 atm; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
2D 1H-13C HSQCsample_1isotropicsample_conditions_1
3D CBCA(CO)NHsample_1isotropicsample_conditions_1
3D C(CO)NHsample_1isotropicsample_conditions_1
3D HNCOsample_1isotropicsample_conditions_1
3D HCCH-TOCSYsample_2isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D 1H-13C NOESYsample_2isotropicsample_conditions_1
3D HCCH-COSYsample_2isotropicsample_conditions_1
2D 1H-15N HSQCsample_5isotropicsample_conditions_2
2D 1H-13C HSQCsample_5isotropicsample_conditions_2
3D CBCA(CO)NHsample_5isotropicsample_conditions_2
3D HNCOsample_5isotropicsample_conditions_2
3D HCCH-COSYsample_5isotropicsample_conditions_2
2D 1H-15N HSQCsample_4isotropicsample_conditions_2

Software:

AZARA v2.7, Boucher - processing

ARIA v2.2, Linge, O'Donoghue and Nilges - chemical shift assignment, refinement

CNS v1.2, Brunger, Adams, Clore, Gros, Nilges and Read - refinement, structure solution

NMRPipe, Goddard - chemical shift assignment, data analysis, peak picking

TALOS, Cornilescu, Delaglio and Bax - refinement

SPARKY v3.113, Goddard - chemical shift assignment, data analysis, peak picking

NMR spectrometers:

  • Bruker Avance 600 MHz

Related Database Links:

BMRB 16482
PDB

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