BMRB Entry 19516

Title:
Solution NMR structure of Dot1L in complex with AF9 (Dot1L-AF9).
Deposition date:
2013-09-20
Original release date:
2015-04-20
Authors:
Kuntimaddi, Aravinda; Bushweller, John
Citation:

Citation: Kuntimaddi, Aravinda; Bushweller, John. "Solution NMR structure of Dot1L in complex with AF9 (Dot1L-AF9)."  .

Assembly members:

Assembly members:
Protein_AF-9, polymer, 69 residues, 8147.291 Da.
Dot1L_Histone-lysine_N-methyltransferase, polymer, 25 residues, 2667.140 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: pETduet-1

Entity Sequences (FASTA):

Entity Sequences (FASTA):
Protein_AF-9: DKAYLDELVELHRRLMTLRE RHILQQIVNLIEETGHFHIT NTTFDFDLCSLDKTTVRKLQ SYLETSGTS
Dot1L_Histone-lysine_N-methyltransferase: TNKLPVSIPLASVVLPSRAE RARST

Data sets:
Data typeCount
13C chemical shifts414
15N chemical shifts89
1H chemical shifts638

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1AF9 protein1
2Dot1L Histone lysine N-methyltransferase2

Entities:

Entity 1, AF9 protein 69 residues - 8147.291 Da.

1   ASPLYSALATYRLEUASPGLULEUVALGLU
2   LEUHISARGARGLEUMETTHRLEUARGGLU
3   ARGHISILELEUGLNGLNILEVALASNLEU
4   ILEGLUGLUTHRGLYHISPHEHISILETHR
5   ASNTHRTHRPHEASPPHEASPLEUCYSSER
6   LEUASPLYSTHRTHRVALARGLYSLEUGLN
7   SERTYRLEUGLUTHRSERGLYTHRSER

Entity 2, Dot1L Histone lysine N-methyltransferase 25 residues - 2667.140 Da.

1   THRASNLYSLEUPROVALSERILEPROLEU
2   ALASERVALVALLEUPROSERARGALAGLU
3   ARGALAARGSERTHR

Samples:

Dot1L-AF9: Protein AF-9, [U-100% 13C; U-100% 15N], 750 uM; Histone-lysine N-methyltransferase, H3 lysine-79 specific, [U-100% 13C; U-100% 15N], 750 uM; Bis-Tris 9.3 mM; MES 15.8 mM; sodium chloride 100 mM; DTT 1 mM; D2O 10%; H2O 95%

Dot1L-AF9_RDC_sample_1: Protein AF-9, [U-100% 13C; U-100% 15N], 750 uM; Histone-lysine N-methyltransferase, H3 lysine-79 specific, [U-100% 13C; U-100% 15N], 750 uM; Bis-Tris 9.3 mM; MES 15.8 mM; sodium chloride 100 mM; DTT 1 mM; D2O 10%; H2O 95%; (3-acrylamidopropyl)-trimethylammonium chloride 3.5%; acrylic acid 3.5%

Dot1L-AF9_RDC_sample_2: Protein AF-9, [U-100% 13C; U-100% 15N], 750 uM; Histone-lysine N-methyltransferase, H3 lysine-79 specific, [U-100% 13C; U-100% 15N], 750 uM; Bis-Tris 9.3 mM; MES 15.8 mM; sodium chloride 100 mM; DTT 1 mM; D2O 10%; H2O 95%; (3-acrylamidopropyl)-trimethylammonium chloride 3.5%; acrylamide 3.5%

Dot1L-AF9_NMR_Sample_Conditions: ionic strength: 100 mM; pH: 6.0; pressure: 1 atm; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
2D 1H-15N HSQCDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
2D 1H-13C HSQCDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D CBCA(CO)NHDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D C(CO)NHDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D HNCODot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D HNCADot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D HNCACBDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D HCCH-TOCSYDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D HNHADot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D 1H-15N NOESYDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D 1H-13C NOESY aliphaticDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D 1H-13C NOESY aromaticDot1L-AF9isotropicDot1L-AF9_NMR_Sample_Conditions
3D HNCO IPAPDot1L-AF9_RDC_sample_1anisotropicDot1L-AF9_NMR_Sample_Conditions
3D HNCO IPAPDot1L-AF9_RDC_sample_2anisotropicDot1L-AF9_NMR_Sample_Conditions

Software:

CYANA v2.1, Guntert, Mumenthaler and Wuthrich - chemical shift assignment

SPARKY, Goddard - peak picking

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

TOPSPIN, Bruker Biospin - collection

X-PLOR NIH v2.34, Schwieters, Kuszewski, Tjandra and Clore - refinement, structure solution

Molmol, Koradi, Billeter and Wuthrich - data analysis

VNMRJ, Varian - collection

NMR spectrometers:

  • Bruker Avance 600 MHz
  • Varian INOVA 600 MHz

Related Database Links:

BMRB 18094
PDB
DBJ BAA04090 BAB31731 BAE27603 BAG35760 BAG72836
EMBL CAA06960 CDQ62412 CDQ68675
GB AAA58361 AAH21420 AAH36089 AAI29090 AAK28536
REF NP_001179478 NP_001273087 NP_001273620 NP_004520 NP_081602
SP A2AM29 P42568
TPG DAA26970
AlphaFold Q96JL1 A2AM29 P42568

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