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BMRB: 15796

2K4D

E2-c-Cbl recognition is necessary but not sufficient for ubiquitination activity

Authors

Huang, A., De Jong, R.N., Hans, W., Sebastiaan, W.G., Marc, T.H.TH., Rolf, B.

Assembly

E2-c-Cbl

Entity

1. E2-c-Cbl (polymer, Thiol state: free and other bound), 83 monomers, 9378.584 Da Detail

GSLQDHIKVT QEQYELYCEM GSTFQLCKIC AENDKDVKIE PCGHLMCTSC LTSWQESEGQ GCPFCRCEIK GTEPIVVDPF DPR

2. ZN (non-polymer), 65.409 × 2 Da

Total weight

9509.402 Da

Max. entity weight

9378.584 Da

Entity Connection

disulfide 8 Detail

ID	Type	Value order	Atom ID 1	Atom ID 2
1	disulfide	sing	1:CYS27:SG	2:ZN1:ZN
2	disulfide	sing	1:CYS30:SG	2:ZN1:ZN
3	disulfide	sing	1:CYS47:SG	2:ZN1:ZN
4	disulfide	sing	1:CYS50:SG	2:ZN1:ZN
5	disulfide	sing	1:CYS42:SG	2:ZN1:ZN
6	disulfide	sing	1:HIS44:ND1	2:ZN1:ZN
7	disulfide	sing	1:CYS62:SG	2:ZN1:ZN
8	disulfide	sing	1:CYS65:SG	2:ZN1:ZN

Source organism

Homo sapiens

Exptl. method

solution NMR

Refine. method

simulated annealing

Data set

assigned_chemical_shifts

Chem. Shift Complete1

Sequence coverage: 97.6 %, Completeness: 96.1 %, Completeness (bb): 97.1 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	96.1 % (912 of 949)	96.8 % (481 of 497)	95.1 % (348 of 366)	96.5 % (83 of 86)
Backbone	97.1 % (474 of 488)	96.4 % (161 of 167)	97.9 % (238 of 243)	96.2 % (75 of 78)
Sidechain	95.5 % (514 of 538)	97.0 % (320 of 330)	93.0 % (186 of 200)	100.0 % (8 of 8)
Aromatic	78.8 % (52 of 66)	93.9 % (31 of 33)	62.5 % (20 of 32)	100.0 % (1 of 1)
Methyl	97.1 % (66 of 68)	94.1 % (32 of 34)	100.0 % (34 of 34)

1. E2-c-Cbl

GSLQDHIKVT QEQYELYCEM GSTFQLCKIC AENDKDVKIE PCGHLMCTSC LTSWQESEGQ GCPFCRCEIK GTEPIVVDPF DPR

Show sample condition

Sample #1

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 7.3

#	Name	Isotope labeling	Concentration
1	entity_1	[U-99% 15N]	1 (±0.2) mM
2	potassium phosphate	natural abundance	50 mM
3	sodium chloride	natural abundance	100 mM
4	zinc chloride	natural abundance	100 uM
5	PMSF	natural abundance	0.2 mM
6	DTT	natural abundance	5 mM

Sample #2

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 7.3

#	Name	Isotope labeling	Concentration
7	entity_1	[U-99% 13C; U-99% 15N]	1 (±0.2) mM
8	potassium phosphate	natural abundance	50 mM
9	sodium chloride	natural abundance	100 mM
10	zinc chloride	natural abundance	100 uM
11	PMSF	natural abundance	0.2 mM
12	DTT	natural abundance	5 mM

Chem. Shift Complete2

Sequence coverage: 31.3 %, Completeness: 56.8 %, Completeness (bb): 59.1 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	56.8 % (1078 of 1898)	56.8 % (565 of 994)	56.4 % (413 of 732)	58.1 % (100 of 172)
Backbone	59.1 % (577 of 976)	59.6 % (199 of 334)	58.8 % (286 of 486)	59.0 % (92 of 156)
Sidechain	55.0 % (592 of 1076)	55.5 % (366 of 660)	54.5 % (218 of 400)	50.0 % (8 of 16)
Aromatic	39.4 % (52 of 132)	47.0 % (31 of 66)	31.3 % (20 of 64)	50.0 % (1 of 2)
Methyl	65.4 % (89 of 136)	66.2 % (45 of 68)	64.7 % (44 of 68)

1. E2-c-Cbl

GSLQDHIKVT QEQYELYCEM GSTFQLCKIC AENDKDVKIE PCGHLMCTSC LTSWQESEGQ GCPFCRCEIK GTEPIVVDPF DPR

Show sample condition

Sample #1

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 7.3

#	Name	Isotope labeling	Concentration
1	entity_1	[U-99% 15N]	1 (±0.2) mM
2	potassium phosphate	natural abundance	50 mM
3	sodium chloride	natural abundance	100 mM
4	zinc chloride	natural abundance	100 uM
5	PMSF	natural abundance	0.2 mM
6	DTT	natural abundance	5 mM

Sample #2

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 7.3

#	Name	Isotope labeling	Concentration
7	entity_1	[U-99% 13C; U-99% 15N]	1 (±0.2) mM
8	potassium phosphate	natural abundance	50 mM
9	sodium chloride	natural abundance	100 mM
10	zinc chloride	natural abundance	100 uM
11	PMSF	natural abundance	0.2 mM
12	DTT	natural abundance	5 mM

Chem. Shift Complete3

Sequence coverage: 31.3 %, Completeness: 45.0 %, Completeness (bb): 48.3 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	45.0 % (1280 of 2847)	44.5 % (663 of 1491)	45.0 % (494 of 1098)	47.7 % (123 of 258)
Backbone	48.3 % (707 of 1464)	48.5 % (243 of 501)	47.9 % (349 of 729)	49.1 % (115 of 234)
Sidechain	42.4 % (684 of 1614)	42.4 % (420 of 990)	42.7 % (256 of 600)	33.3 % (8 of 24)
Aromatic	28.8 % (57 of 198)	36.4 % (36 of 99)	20.8 % (20 of 96)	33.3 % (1 of 3)
Methyl	48.0 % (98 of 204)	47.1 % (48 of 102)	49.0 % (50 of 102)

1. E2-c-Cbl

GSLQDHIKVT QEQYELYCEM GSTFQLCKIC AENDKDVKIE PCGHLMCTSC LTSWQESEGQ GCPFCRCEIK GTEPIVVDPF DPR

Show sample condition

Sample #1

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 7.3

#	Name	Isotope labeling	Concentration
1	entity_1	[U-99% 15N]	1 (±0.2) mM
2	potassium phosphate	natural abundance	50 mM
3	sodium chloride	natural abundance	100 mM
4	zinc chloride	natural abundance	100 uM
5	PMSF	natural abundance	0.2 mM
6	DTT	natural abundance	5 mM

Sample #2

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 7.3

#	Name	Isotope labeling	Concentration
7	entity_1	[U-99% 13C; U-99% 15N]	1 (±0.2) mM
8	potassium phosphate	natural abundance	50 mM
9	sodium chloride	natural abundance	100 mM
10	zinc chloride	natural abundance	100 uM
11	PMSF	natural abundance	0.2 mM
12	DTT	natural abundance	5 mM

Protein Blocks Logo

Calculated from 20 models in PDB: 2K4D, Strand ID: A Detail

Release date

2008-11-13

Citation

E2-c-Cbl recognition is necessary but not sufficient for ubiquitination activity
Huang, A., De Jong, R.N., Wienk, H., Winkler, S., Timmers, M., Boelens, R.
J. Mol. Biol. (2009), 385, 507-519, PubMed 18996392 , DOI 10.1016/j.jmb.2008.10.044 , Abstract

Related entities 1. E2-c-Cbl, : 1 : 10 : 50 entities Detail

Interaction partners 1. E2-c-Cbl, : 27 interactors Detail

More details for 27 interactors identified by 16 citations

Experiments performed 16 experiments Detail

1 2D 1H-15N HSQC