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SOLUTION STRUCTURE OF LACTOBACILLUS CASEI DIHYDROFOLATE REDUCTASE APO-FORM, 25 CONFORMERS
Authors
Polshakov, V.I., Birdsall, B., Feeney, J.
Assembly
apo DHFR
Entity
1. apo DHFR (polymer, Thiol state: not present), 162 monomers, 18307.38 Da Detail

TAFLWAQDRD GLIGKDGHLP WHLPDDLHYF RAQTVGKIMV VGRRTYESFP KRPLPERTNV VLTHQEDYQA QGAVVVHDVA AVFAYAKQHP DQELVIAGGA QIFTAFKDDV DTLLVTRLAG SFEGDTKMIP LNWDDFTKVS SRTVEDTNPA LTHTYEVWQK KA


Formula weight
18307.38 Da
Source organism
Lacticaseibacillus casei
Exptl. method
solution NMR
Refine. method
simulated annealing
Data set
assigned_chemical_shifts, H_exch_protection_factors
Chem. Shift Complete
Sequence coverage: 96.9 %, Completeness: 85.1 %, Completeness (bb): 88.5 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All85.1 % (1608 of 1890)88.2 % (856 of 971)80.0 % (599 of 749)90.0 % (153 of 170)
Backbone88.5 % (846 of 956)90.5 % (295 of 326)85.9 % (409 of 476)92.2 % (142 of 154)
Sidechain82.7 % (898 of 1086)87.0 % (561 of 645)76.7 % (326 of 425)68.8 % (11 of 16)
Aromatic64.3 % (126 of 196)77.6 % (76 of 98)53.2 % (50 of 94) 0.0 % (0 of 4)
Methyl91.8 % (178 of 194)92.8 % (90 of 97)90.7 % (88 of 97)

1. DHFR

TAFLWAQDRD GLIGKDGHLP WHLPDDLHYF RAQTVGKIMV VGRRTYESFP KRPLPERTNV VLTHQEDYQA QGAVVVHDVA AVFAYAKQHP DQELVIAGGA QIFTAFKDDV DTLLVTRLAG SFEGDTKMIP LNWDDFTKVS SRTVEDTNPA LTHTYEVWQK KA

Sample #1

Solvent system 100% D2O, Pressure 1 atm, Temperature 288 K, pH 6.5


#NameIsotope labelingTypeConcentration
1DHFRnatural abundance2 mM
2potassium chloridenatural abundance100 mM
3potassium phosphatenatural abundance50 mM
4sodium azidenatural abundance0.1 %
5D2Onatural abundance100 %
Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 288 K, pH 6.5


#NameIsotope labelingTypeConcentration
6DHFR[U-99% 13C; U-99% 15N]1 mM
7potassium chloridenatural abundance100 mM
8potassium phosphatenatural abundance50 mM
9sodium azidenatural abundance0.1 %
10D2Onatural abundance10 %
11H2Onatural abundance90 %
Sample #3

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 288 K, pH 6.5


#NameIsotope labelingTypeConcentration
12DHFR[U-99% 15N]1 mM
13potassium chloridenatural abundance100 mM
14potassium phosphatenatural abundance50 mM
15sodium azidenatural abundance0.1 %
16D2Onatural abundance5 %
17H2Onatural abundance95 %
Sample #4

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 288 K, pH 6.5


#NameIsotope labelingTypeConcentration
18DHFR[U-99% 15N]1 mM
19potassium chloridenatural abundance100 mM
20potassium phosphatenatural abundance50 mM
21sodium azidenatural abundance0.1 %
22DMPC/DHPCnatural abundance5 %
23D2Onatural abundance5 %
24H2Onatural abundance95 %

LACS Plot; CA
Referencing offset: 0.37 ppm, Outliers: 1 Detail
LACS Plot; CB
Referencing offset: 0.37 ppm, Outliers: 1 Detail
LACS Plot; HA
Referencing offset: -0.21 ppm, Outliers: 1 Detail
LACS Plot; CO
Referencing offset: 0.77 ppm, Outliers: 1 Detail
Protein Blocks Logo
Calculated from 25 models in PDB: 2L28, Strand ID: A Detail


1H exch. protect.
54 values in 1 lists
Calculation method entry_citation, Pressure 1 atm, Temperature 288 K, pH 6.5 Detail
Release date
2011-05-17
Citation
NMR structures of apo L. casei dihydrofolate reductase and its complexes with trimethoprim and NADPH: contributions to positive cooperative binding from ligand-induced refolding, conformational changes, and interligand hydrophobic interactions
Feeney, J., Birdsall, B., Kovalevskaya, N.V., Smurnyy, Y.D., Navarro Peran, E.M., Polshakov, V.I.
Biochemistry (2011), 50, 3609-3620, PubMed 21410224 , DOI 10.1021/bi200067t ,
Related entities 1. apo DHFR, : 1 : 9 : 137 entities Detail
Experiments performed 16 experiments Detail
nullKeywords antifolates, co-operative binding