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BMRB: 5299

1KLQ

The Mad2 Spindle Checkpoint Protein Undergoes Similar Major Conformational Changes upon Binding to Either Mad1 or Cdc20

Authors

Luo, X., Tang, Z., Rizo, J., Yu, H.

Assembly

MITOTIC SPINDLE ASSEMBLY CHECKPOINT PROTEIN MAD2A/PEPTIDE

Entity

1. mitotic checkpoint protein Mad2 (polymer, Thiol state: all free), 197 monomers, 22539.50 Da Detail

GSITLRGSAE IVAEFFSFGI NSILYQRGIY PSETFTRVQK YGLTLLVTTD LELIKYLNNV VEQLKDWLYK CSVQKLVVVI SNIESGEVLE RWQFDIECDK TAKDDSAPRE KSQKAIQDEI RSVIRQITAT VTFLPLLEVS CSFDLLIYTD KDLVVPEKWE ESGPQFITNS EEVRLRSFTT TIHKVNSMVA YKIPVND

2. Mad2-binding peptide (polymer, Thiol state: not present), 12 monomers, 1450.595 Da Detail

SWYSYPPPQR AV

Total weight

23990.096 Da

Max. entity weight

22539.5 Da

Source organism

Homo sapiens

Exptl. method

NMR

Refine. method

torsion angle dynamics

Data set

assigned_chemical_shifts

Chem. Shift Complete1

Sequence coverage: 93.8 %, Completeness: 74.1 %, Completeness (bb): 91.5 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	74.1 % (1864 of 2515)	70.4 % (917 of 1302)	74.6 % (740 of 992)	93.7 % (207 of 221)
Backbone	91.5 % (1131 of 1236)	93.0 % (387 of 416)	89.4 % (554 of 620)	95.0 % (190 of 200)
Sidechain	61.4 % (910 of 1481)	59.7 % (529 of 886)	63.4 % (364 of 574)	81.0 % (17 of 21)
Aromatic	47.7 % (102 of 214)	78.5 % (84 of 107)	14.6 % (15 of 103)	75.0 % (3 of 4)
Methyl	89.3 % (241 of 270)	91.1 % (123 of 135)	87.4 % (118 of 135)

1. mitotic checkpoint protein Mad2

2. Mad2-binding peptide

SWYSYPPPQR AV

Show sample condition

Sample

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Chem. Shift Complete2

Sequence coverage: 32.5 %, Completeness: 46.7 %, Completeness (bb): 57.3 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	46.7 % (2351 of 5030)	44.8 % (1166 of 2604)	46.3 % (918 of 1984)	60.4 % (267 of 442)
Backbone	57.3 % (1417 of 2472)	59.1 % (492 of 832)	54.8 % (679 of 1240)	61.5 % (246 of 400)
Sidechain	38.7 % (1146 of 2962)	37.8 % (670 of 1772)	39.7 % (456 of 1148)	47.6 % (20 of 42)
Aromatic	32.0 % (137 of 428)	48.1 % (103 of 214)	14.6 % (30 of 206)	50.0 % (4 of 8)
Methyl	51.7 % (279 of 540)	53.7 % (145 of 270)	49.6 % (134 of 270)

1. mitotic checkpoint protein Mad2

2. Mad2-binding peptide

SWYSYPPPQR AV

Show sample condition

Sample

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

Protein Blocks Logo

Calculated from 1 models in PDB: 1KLQ, Strand ID: A, B Detail

Release date

2002-03-21

Citation

The Mad2 spindle checkpoint protein undergoes similar major conformational changes upon binding to either Mad1 or Cdc20
Luo, X., Tang, Z., Rizo, J., Yu, H.
Mol. Cell (2002), 9, 59-71, PubMed 11804586 , DOI 10.2210/pdb1klq/pdb , Abstract

Related entities 1. mitotic checkpoint protein Mad2, : 5 : 19 entities Detail

Related entities 2. Mad2-binding peptide, : 1 : 2 : 1 entities Detail

Experiments performed 8 experiments Detail

1 HNCA

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

2 3D H(CC)(CO)NH

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

3 3D 15N-separated NOESY

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

4 3D 13C-separated NOESY

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

5 HN(CO)CA

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

6 3D (H)C(C)(CO)NH

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

7 HN(COCA)CB

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

8 HN(CA)CB

Instrument

Model Varian INOVA (600 MHz)

Sample #1

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
1	mitotic checkpoint protein Mad2	[U-15N]	0.8 mM
2	Mad2-binding peptide		1 mM
3	phosphate buffer		50 mM
4	KCl		0.3 M
5	DTT		1 mM
6	D2O		10 %
7	H2O		90 %

Sample #2

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
8	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-2H]	0.8 mM
9	Mad2-binding peptide		1 mM
10	phosphate buffer		50 mM
11	KCl		0.3 M
12	DTT		1 mM
13	D2O		10 %
14	H2O		90 %

Sample #3

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
15	mitotic checkpoint protein Mad2	[U-15N; U-13C]	0.8 mM
16	Mad2-binding peptide		1 mM
17	phosphate buffer		50 mM
18	KCl		0.3 M
19	DTT		1 mM
20	D2O		10 %
21	H2O		90 %

Sample #4

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
22	mitotic checkpoint protein Mad2	[U-15N; U-13C; U-60% 2H]	0.8 mM
23	Mad2-binding peptide		1 mM
24	phosphate buffer		50 mM
25	KCl		0.3 M
26	DTT		1 mM
27	D2O		10 %
28	H2O		90 %

Sample #5

Pressure 1 atm, Temperature 303 (±0.1) K, pH 7.4 (±0.1)

#	Name	Isotope labeling	Concentration
29	mitotic checkpoint protein Mad2		1 mM
30	Mad2-binding peptide	[U-13C; U-15N]	0.8 mM
31	phosphate buffer		50 mM
32	KCl		0.3 M
33	DTT		1 mM
34	D2O		10 %
35	H2O		90 %

nullKeywords protein-peptide complex, MAD2 FAMILY

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Found 1 Document (1.597 seconds)

BMRB: 5299

Sample

Sample

Related entities 1. mitotic checkpoint protein Mad2, : 5 : 19 entities Detail

Related entities 2. Mad2-binding peptide, : 1 : 2 : 1 entities Detail

Experiments performed 8 experiments Detail

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