Search

1H, 15N and 13C resonance assignments of human dihydrofolate reductase in its complex with trimethoprim and NADPH
Authors
Kovalevskaya, N.V., Polshakov, V.I., Birdsall, B., Bradbury, A.F., Frenkiel, T.A., Feeney, J.
Assembly
ternary complex of human DHFR with trimethoprim and NADPH
Entity
1. Dihydrofolate reductase (polymer, Thiol state: all free), 186 monomers, 21321.27 Da Detail

VGSLNCIVAV SQNMGIGKNG DLPWPPLRNE FRYFQRMTTT SSVEGKQNLV IMGKKTWFSI PEKNRPLKGR INLVLSRELK EPPQGAHFLS RSLDDALKLT EQPELANKVD MVWIVGGSSV YKEAMNHPGH LKLFVTRIMQ DFESDTFFPE IDLEKYKLLP EYPGVLSDVQ EEKGIKYKFE VYEKND


2. NAP (non-polymer), 743.405 Da
3. TOP (non-polymer), 290.318 Da
Total weight
22354.992 Da
Max. entity weight
21321.27 Da
Source organism
Homo sapiens
Exptl. method
NMR
Refine. method
simulated annealing, molecular dynamics
Data set
assigned_chemical_shifts
Chem. Shift Complete1
Sequence coverage: 100.0 %, Completeness: 80.9 %, Completeness (bb): 95.2 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All80.9 % (1840 of 2275)79.1 % (944 of 1193)79.2 % (697 of 880)98.5 % (199 of 202)
Backbone95.2 % (1027 of 1079)95.6 % (344 of 360)93.4 % (509 of 545)100.0 % (174 of 174)
Sidechain71.4 % (977 of 1369)72.0 % (600 of 833)69.3 % (352 of 508)89.3 % (25 of 28)
Aromatic58.1 % (108 of 186)64.5 % (60 of 93)50.0 % (45 of 90)100.0 % (3 of 3)
Methyl88.0 % (169 of 192)97.9 % (94 of 96)78.1 % (75 of 96)

1. Dihydrofolate reductase

VGSLNCIVAV SQNMGIGKNG DLPWPPLRNE FRYFQRMTTT SSVEGKQNLV IMGKKTWFSI PEKNRPLKGR INLVLSRELK EPPQGAHFLS RSLDDALKLT EQPELANKVD MVWIVGGSSV YKEAMNHPGH LKLFVTRIMQ DFESDTFFPE IDLEKYKLLP EYPGVLSDVQ EEKGIKYKFE VYEKND

Sample #1

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
1Dihydrofolate reductase0.5 ~ 1.2 mM
2TRIMETHOPRIM0.5 ~ 1.2 mM
3NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 ~ 1.2 mM
4potassium phosphate50 mM
5potassium chloride100 mM
Sample #2

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
6Dihydrofolate reductase[U-98% 13C; U-98% 15N]0.5 ~ 1.0 mM
7TRIMETHOPRIM0.5 ~ 1.0 mM
8NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 ~ 1.0 mM
9potassium phosphate50 mM
10potassium chloride100 mM
Sample #3

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
11Dihydrofolate reductase[U-98% 15N]0.75 ~ 1.2 mM
12TRIMETHOPRIM0.75 ~ 1.2 mM
13NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.75 ~ 1.2 mM
14potassium phosphate50 mM
15potassium chloride100 mM

Chem. Shift Complete2
Sequence coverage: 15.1 %, Completeness: 45.8 %, Completeness (bb): 54.4 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All45.8 % (2083 of 4550)44.7 % (1066 of 2386)44.8 % (789 of 1760)56.4 % (228 of 404)
Backbone54.4 % (1174 of 2158)54.7 % (394 of 720)53.0 % (578 of 1090)58.0 % (202 of 348)
Sidechain40.0 % (1094 of 2738)40.3 % (672 of 1666)39.0 % (396 of 1016)46.4 % (26 of 56)
Aromatic29.0 % (108 of 372)32.3 % (60 of 186)25.0 % (45 of 180)50.0 % (3 of 6)
Methyl49.7 % (191 of 384)54.7 % (105 of 192)44.8 % (86 of 192)

1. Dihydrofolate reductase

VGSLNCIVAV SQNMGIGKNG DLPWPPLRNE FRYFQRMTTT SSVEGKQNLV IMGKKTWFSI PEKNRPLKGR INLVLSRELK EPPQGAHFLS RSLDDALKLT EQPELANKVD MVWIVGGSSV YKEAMNHPGH LKLFVTRIMQ DFESDTFFPE IDLEKYKLLP EYPGVLSDVQ EEKGIKYKFE VYEKND

Sample #1

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
1Dihydrofolate reductase0.5 ~ 1.2 mM
2TRIMETHOPRIM0.5 ~ 1.2 mM
3NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 ~ 1.2 mM
4potassium phosphate50 mM
5potassium chloride100 mM
Sample #2

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
6Dihydrofolate reductase[U-98% 13C; U-98% 15N]0.5 ~ 1.0 mM
7TRIMETHOPRIM0.5 ~ 1.0 mM
8NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 ~ 1.0 mM
9potassium phosphate50 mM
10potassium chloride100 mM
Sample #3

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
11Dihydrofolate reductase[U-98% 15N]0.75 ~ 1.2 mM
12TRIMETHOPRIM0.75 ~ 1.2 mM
13NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.75 ~ 1.2 mM
14potassium phosphate50 mM
15potassium chloride100 mM

Chem. Shift Complete3
Sequence coverage: 23.7 %, Completeness: 35.6 %, Completeness (bb): 42.6 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All35.6 % (2427 of 6825)34.5 % (1234 of 3579)35.0 % (925 of 2640)44.2 % (268 of 606)
Backbone42.6 % (1380 of 3237)42.3 % (457 of 1080)41.7 % (682 of 1635)46.2 % (241 of 522)
Sidechain30.8 % (1267 of 4107)31.1 % (777 of 2499)30.4 % (463 of 1524)32.1 % (27 of 84)
Aromatic20.6 % (115 of 558)24.0 % (67 of 279)16.7 % (45 of 270)33.3 % (3 of 9)
Methyl37.0 % (213 of 576)40.6 % (117 of 288)33.3 % (96 of 288)

1. Dihydrofolate reductase

VGSLNCIVAV SQNMGIGKNG DLPWPPLRNE FRYFQRMTTT SSVEGKQNLV IMGKKTWFSI PEKNRPLKGR INLVLSRELK EPPQGAHFLS RSLDDALKLT EQPELANKVD MVWIVGGSSV YKEAMNHPGH LKLFVTRIMQ DFESDTFFPE IDLEKYKLLP EYPGVLSDVQ EEKGIKYKFE VYEKND

Sample #1

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
1Dihydrofolate reductase0.5 ~ 1.2 mM
2TRIMETHOPRIM0.5 ~ 1.2 mM
3NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 ~ 1.2 mM
4potassium phosphate50 mM
5potassium chloride100 mM
Sample #2

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
6Dihydrofolate reductase[U-98% 13C; U-98% 15N]0.5 ~ 1.0 mM
7TRIMETHOPRIM0.5 ~ 1.0 mM
8NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 ~ 1.0 mM
9potassium phosphate50 mM
10potassium chloride100 mM
Sample #3

Temperature 288 (±1) K, pH 6.5 (±0.1)


#NameIsotope labelingTypeConcentration
11Dihydrofolate reductase[U-98% 15N]0.75 ~ 1.2 mM
12TRIMETHOPRIM0.75 ~ 1.2 mM
13NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.75 ~ 1.2 mM
14potassium phosphate50 mM
15potassium chloride100 mM

LACS Plot; CA
Referencing offset: -0.09 ppm, Outliers: 2 Detail
LACS Plot; CB
Referencing offset: -0.09 ppm, Outliers: 2 Detail
LACS Plot; HA
Referencing offset: -0.06 ppm, Outliers: 1 Detail
LACS Plot; CO
Referencing offset: 0.23 ppm, Outliers: 4 Detail
Protein Blocks Logo
Calculated from 25 models in PDB: 1YHO, Strand ID: A Detail


Release date
2006-01-18
Citation
Solution structure of human dihydrofolate reductase in its complex with trimethoprim and NADPH
Kovalevskaya, N.V., Smurnyy, Y.D., Polshakov, V.I., Birdsall, B., Bradbury, A.F., Frenkiel, T.A., Feeney, J.
J. Biomol. NMR (2005), 33, 69-72, PubMed 16222560 , DOI 10.1007/s10858-005-1475-z ,
Related entities 1. Dihydrofolate reductase, : 1 : 61 : 4 : 9 : 91 entities Detail
Interaction partners 1. Dihydrofolate reductase, : 4 interactors Detail
Keywords Dihydrofolate reductase, Trimethoprim, Protein-ligand interactions, Co-operative binding, NMR resonance assignments