BMRB: 51145

K63-linked diubiquitin with 15N-labeled proximal ubiquitin interacting with ASCC2 CUE domain

Authors

Lombardi, P., Majumdar, A., Wolberger, C.

Assembly

K63Ub2:ASCC2 CUE

Entity

1. K63Ub2:ASCC2 CUE, entity 1 (polymer, Thiol state: not present), 153 monomers, 17282.56 Da Detail

MQIFVKTLTG KTITLEVEPS DTIENVKAKI QDKEGIPPDQ QRLIFAGRQL EDGRTLSDYN IQRESTLHLV LRLRGGMQIF VKTLTGKTIT LEVEPSDTIE NVKAKIQDKE GIPPDQQRLI FAGKQLEDGR TLSDYNIQKE STLHLVLRLR GGD

2. K63Ub2:ASCC2 CUE, entity 2 (polymer, Thiol state: all free), 55 monomers, 6331.075 Da Detail

ELDSLISQVK DLLPDLGEGF ILACLEYYHY DPEQVINNIL EERLAPTLSQ LDRNL

Total weight

23613.637 Da

Max. entity weight

17282.56 Da

Source organism

Homo sapiens

Exptl. method

solution NMR

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 36.5 %, Completeness: 10.3 %, Completeness (bb): 25.0 % Detail

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Polymer type: polypeptide(L)

	Total	1H	15N
All	10.3 % (158 of 1531)	6.4 % (83 of 1298)	32.2 % (75 of 233)
Backbone	25.0 % (155 of 620)	19.0 % (80 of 421)	37.7 % (75 of 199)
Sidechain	0.3 % (3 of 911)	0.3 % (3 of 877)	0.0 % (0 of 34)
Aromatic	0.0 % (0 of 51)	0.0 % (0 of 51)
Methyl	0.0 % (0 of 137)	0.0 % (0 of 137)

1. entity 1

MQIFVKTLTG KTITLEVEPS DTIENVKAKI QDKEGIPPDQ QRLIFAGRQL EDGRTLSDYN IQRESTLHLV LRLRGGMQIF VKTLTGKTIT LEVEPSDTIE NVKAKIQDKE GIPPDQQRLI FAGKQLEDGR TLSDYNIQKE STLHLVLRLR GGD

2. entity 2

ELDSLISQVK DLLPDLGEGF ILACLEYYHY DPEQVINNIL EERLAPTLSQ LDRNL

Show sample condition

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 293 K, pH 7.0, Details 100 uM K63-linked diubiquitin (15N-labeled proximal ubiquitin) titrated with 0, 30, 100, 200, and 500 uM ASCC2 CUE domain.

#	Name	Isotope labeling	Type	Concentration
1	ASCC2 CUE domain	natural abundance		500 uM
2	K63-linked diubiquitin	[U-15N]		100 uM
3	Tris	natural abundance		20 mM
4	NaCl	natural abundance		100 mM
5	TCEP	natural abundance		200 mM

Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 293 K, pH 7.0, Details 100 uM K63-linked diubiquitin (15N-labeled proximal ubiquitin) titrated with 0, 30, 100, 200, and 500 uM ASCC2 CUE domain.

#	Name	Isotope labeling	Type	Concentration
6	ASCC2 CUE domain	natural abundance		200 uM
7	K63-linked diubiquitin	[U-15N]		100 uM
8	Tris	natural abundance		20 mM
9	NaCl	natural abundance		100 mM
10	TCEP	natural abundance		200 mM

Sample #3

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 293 K, pH 7.0, Details 100 uM K63-linked diubiquitin (15N-labeled proximal ubiquitin) titrated with 0, 30, 100, 200, and 500 uM ASCC2 CUE domain.

#	Name	Isotope labeling	Type	Concentration
11	ASCC2 CUE domain	natural abundance		100 uM
12	K63-linked diubiquitin	[U-15N]		100 uM
13	Tris	natural abundance		20 mM
14	NaCl	natural abundance		100 mM
15	TCEP	natural abundance		200 mM

Sample #4

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 293 K, pH 7.0, Details 100 uM K63-linked diubiquitin (15N-labeled proximal ubiquitin) titrated with 0, 30, 100, 200, and 500 uM ASCC2 CUE domain.

#	Name	Isotope labeling	Type	Concentration
16	ASCC2 CUE domain	natural abundance		30 uM
17	K63-linked diubiquitin	[U-15N]		100 uM
18	Tris	natural abundance		20 mM
19	NaCl	natural abundance		100 mM
20	TCEP	natural abundance		200 mM

Sample #5

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 293 K, pH 7.0, Details 100 uM K63-linked diubiquitin (15N-labeled proximal ubiquitin) titrated with 0, 30, 100, 200, and 500 uM ASCC2 CUE domain.

#	Name	Isotope labeling	Type	Concentration
21	K63-linked diubiquitin	[U-15N]		100 uM
22	Tris	natural abundance		20 mM
23	NaCl	natural abundance		100 mM
24	TCEP	natural abundance		200 mM

Hide sample codition

Release date

2021-10-17

Citation

The ASCC2 CUE domain in the ALKBH3-ASCC DNA repair complex recognizes adjacent ubiquitins in K63-linked polyubiquitin
Lombardi, P.M., Haile, S., Rusanov, T., Rodell, R., Anoh, R., Baer, J.G., Burke, K.A., Gray, L.N., Hacker, A.R., Kebreau, K.R., Ngandu, C.K., Orland, H.A., Osei-Asante, E., Schmelyun, D.P., Shorb, D.E., Syed, S.H., Veilleux, J.M., Majumdar, A., Mosammaparast, N., Wolberger, C.
J. Biol. Chem. (2022), 298, 101545-101545, PubMed 34971705 , DOI 10.1016/j.jbc.2021.101545 , Abstract

Entries sharing articles BMRB: 3 entries Detail

Related entities 1. K63Ub2:ASCC2 CUE, entity 1, : 221 entities Detail

Related entities 2. K63Ub2:ASCC2 CUE, entity 2, : 1 : 2 : 12 entities Detail

Interaction partners 2. K63Ub2:ASCC2 CUE, entity 2, : 49 interactors Detail

Experiments performed 1 experiments Detail

Chemical shift validation 4 contents Detail