Evidence: NMR Distance Restraints, We observe intermolecular NOE contacts between the protein chains, homology, TAZ2 binds many different activation domains in a 1:1 manner, isothermal titration ...Evidence: NMR Distance Restraints, We observe intermolecular NOE contacts between the protein chains, homology, TAZ2 binds many different activation domains in a 1:1 manner, isothermal titration calorimetry, We observe 1:1 binding via ITC
Type
Name
Symmetry operation
Number
identity operation
1_555
1
Buried area
1170 Å2
ΔGint
-2 kcal/mol
Surface area
9080 Å2
NMR ensembles
Data
Criteria
Number of conformers (submitted / calculated)
20 / 100
structures with the lowest energy
Representative
Model #1
lowest energy
-
Components
#1: Protein/peptide
Microphthalmia-associatedtranscriptionfactor / Class E basic helix-loop-helix protein 32 / bHLHe32
Mass: 3720.123 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: MITF, BHLHE32 / Production host: Escherichia coli (E. coli) / References: UniProt: O75030
#2: Protein
Histoneacetyltransferasep300 / p300 HAT / E1A-associated protein p300 / Histone butyryltransferase p300 / Histone ...p300 HAT / E1A-associated protein p300 / Histone butyryltransferase p300 / Histone crotonyltransferase p300 / Protein 2-hydroxyisobutyryltransferase p300 / Protein lactyltransferas p300 / Protein propionyltransferase p300
Mass: 10107.916 Da / Num. of mol.: 1 / Mutation: C1738A C1746A C1789A C1790A Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: EP300, P300 / Production host: Escherichia coli (E. coli) References: UniProt: Q09472, histone acetyltransferase, Transferases; Acyltransferases; Transferring groups other than aminoacyl groups
Mass: 65.409 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: Zn
Has ligand of interest
N
-
Experimental details
-
Experiment
Experiment
Method: SOLUTION NMR
NMR experiment
Conditions-ID
Experiment-ID
Solution-ID
Sample state
Spectrometer-ID
Type
1
1
1
isotropic
1
2D 1H-15N HSQC
1
2
1
isotropic
1
2D 1H-13C HSQC
1
4
1
isotropic
1
3D HNCO
1
11
1
isotropic
1
3D HCACO
1
10
1
isotropic
1
3D HN(CA)CB
1
9
1
isotropic
1
3DCBCA(CO)NH
1
8
1
isotropic
1
3DC(CO)NH
1
7
1
isotropic
1
3DH(CCO)NH
1
6
1
isotropic
1
3D (H)CCH-COSY
1
5
1
isotropic
1
3D (H)CCH-TOCSY
1
3
1
isotropic
1
3DHBHA(CO)NH
1
16
1
isotropic
1
3D 1H-13C NOESY aliphatic
1
15
1
isotropic
1
3D 1H-15N NOESY
1
14
2
isotropic
1
2D 1H-13C HSQC
1
13
2
isotropic
1
2D 1H-15N HSQC
1
12
2
isotropic
1
3D HNCO
1
26
2
isotropic
1
3D HCACO
1
25
2
isotropic
1
3D HN(CA)CB
1
24
2
isotropic
1
3DCBCA(CO)NH
1
23
2
isotropic
1
3DC(CO)NH
1
22
2
isotropic
1
3DH(CCO)NH
1
21
2
isotropic
1
3D (H)CCH-COSY
1
20
2
isotropic
1
3D (H)CCH-TOCSY
1
19
2
isotropic
1
3DHBHA(CO)NH
1
18
2
isotropic
1
3D 1H-13C NOESY aliphatic
1
17
2
isotropic
1
3D 1H-13C NOESY aromatic
1
29
2
isotropic
1
3D 1H-15N NOESY
-
Sample preparation
Details
Type
Solution-ID
Contents
Label
Solvent system
solution
1
20 mM 2-(N-morpholino)ethanesulfonic acid (MES), 5 mM beta mercaptoethanol, 10 uM Zinc chloride, 1 mM [U-13C; U-15N] Microphthalmia-associated transcription factor, 1.2 mM TAZ2 domain of p300, 95% H2O/5% D2O
MITF-labelled complex
95% H2O/5% D2O
solution
2
20 mM 2-(N-morpholino)ethanesulfonic acid (MES), 5 mM beta mercaptoethanol, 10 mM Zinc chloride, 1.15 mM Microphthalmia-associated transcription factor, 0.95 mM [U-13C; U-15N] TAZ2 domain of p300, 95% H2O/5% D2O
Taz2-labelled complex
95% H2O/5% D2O
Sample
Conc. (mg/ml)
Component
Isotopic labeling
Solution-ID
20mM
2-(N-morpholino)ethanesulfonic acid (MES)
naturalabundance
1
5mM
betamercaptoethanol
naturalabundance
1
10uM
Zincchloride
naturalabundance
1
1mM
Microphthalmia-associated transcription factor
[U-13C; U-15N]
1
1.2mM
TAZ2domainofp300
naturalabundance
1
20mM
2-(N-morpholino)ethanesulfonic acid (MES)
naturalabundance
2
5mM
betamercaptoethanol
naturalabundance
2
10mM
Zincchloride
naturalabundance
2
1.15mM
Microphthalmia-associated transcription factor
naturalabundance
2
0.95mM
TAZ2domainofp300
[U-13C; U-15N]
2
Sample conditions
Ionic strength: 20 mM / Label: condition 1 / pH: 6.0 / Pressure: 1 atm / Temperature: 308 K
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