BMRB

Biological Magnetic Resonance Data Bank


A Repository for Data from NMR Spectroscopy on Proteins, Peptides, Nucleic Acids, and other Biomolecules
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BMRB Entry 5981

Title: 1H, 15N and 13C resonance assignments of human dihydrofolate reductase in its complex with trimethoprim and NADPH   PubMed: 16222560

Authors: Kovalevskaya, Nadezhda; Polshakov, Vladimir; Birdsall, Berry; Bradbury, Alan; Frenkiel, Thomas; Feeney, James

Citation: Kovalevskaya, Nadezhda; Smurnyy, Yegor; Polshakov, Vladimir; Birdsall, Berry; Bradbury, Alan; Frenkiel, Thomas; Feeney, James. "Solution structure of human dihydrofolate reductase in its complex with trimethoprim and NADPH "  J. Biomol. NMR 33, 69-72 (2005).

Assembly members:
Dihydrofolate reductase, polymer, 186 residues, 21300 Da.
NAP, non-polymer, 743.405 Da.
TOP, non-polymer, 290.318 Da.

Natural source:   Common Name: Human   Taxonomy ID: 9606   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Homo sapiens

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli

Entity Sequences (FASTA):
Dihydrofolate reductase: VGSLNCIVAVSQNMGIGKNG DLPWPPLRNEFRYFQRMTTT SSVEGKQNLVIMGKKTWFSI PEKNRPLKGRINLVLSRELK EPPQGAHFLSRSLDDALKLT EQPELANKVDMVWIVGGSSV YKEAMNHPGHLKLFVTRIMQ DFESDTFFPEIDLEKYKLLP EYPGVLSDVQEEKGIKYKFE VYEKND

Data sets:
Data typeCount
1H chemical shifts1146
13C chemical shifts706
15N chemical shifts204

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
  • Hide all

Assembly:

Entity Assembly IDEntity NameEntity ID
1dihydrofolate reductase1
2trimethoprim3
3NADPH2

Entities:

Entity 1, dihydrofolate reductase 186 residues - 21300 Da.

1   VALGLYSERLEUASNCYSILEVALALAVAL
2   SERGLNASNMETGLYILEGLYLYSASNGLY
3   ASPLEUPROTRPPROPROLEUARGASNGLU
4   PHEARGTYRPHEGLNARGMETTHRTHRTHR
5   SERSERVALGLUGLYLYSGLNASNLEUVAL
6   ILEMETGLYLYSLYSTHRTRPPHESERILE
7   PROGLULYSASNARGPROLEULYSGLYARG
8   ILEASNLEUVALLEUSERARGGLULEULYS
9   GLUPROPROGLNGLYALAHISPHELEUSER
10   ARGSERLEUASPASPALALEULYSLEUTHR
11   GLUGLNPROGLULEUALAASNLYSVALASP
12   METVALTRPILEVALGLYGLYSERSERVAL
13   TYRLYSGLUALAMETASNHISPROGLYHIS
14   LEULYSLEUPHEVALTHRARGILEMETGLN
15   ASPPHEGLUSERASPTHRPHEPHEPROGLU
16   ILEASPLEUGLULYSTYRLYSLEULEUPRO
17   GLUTYRPROGLYVALLEUSERASPVALGLN
18   GLUGLULYSGLYILELYSTYRLYSPHEGLU
19   VALTYRGLULYSASNASP

Entity 3, trimethoprim - C14 H18 N4 O3 - 290.318 Da.

1   TOP

Entity 2, NADPH - C21 H28 N7 O17 P3 - 743.405 Da.

1   NAP

Samples:

unl-DHFR-TMP-NADPH: Dihydrofolate reductase0.5 – 1.2 mM; TRIMETHOPRIM0.5 – 1.2 mM; NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 – 1.2 mM; potassium phosphate 50 mM; potassium chloride 100 mM

13C-15N-DHFR-TMP-NADPH: Dihydrofolate reductase, [U-98% 13C; U-98% 15N], 0.5 – 1.0 mM; TRIMETHOPRIM0.5 – 1.0 mM; NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.5 – 1.0 mM; potassium phosphate 50 mM; potassium chloride 100 mM

15N-DHFR-TMP-NADPH: Dihydrofolate reductase, [U-98% 15N], 0.75 – 1.2 mM; TRIMETHOPRIM0.75 – 1.2 mM; NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE0.75 – 1.2 mM; potassium phosphate 50 mM; potassium chloride 100 mM

condition_1: pH: 6.5; temperature: 288 K

Experiments:

NameSampleSample stateSample conditions
2D DQF-COSYnot availablenot availablecondition_1
2D NOESYnot availablenot availablecondition_1
2D 1H-15N HSQCnot availablenot availablecondition_1
2D 1H-13C HSQCnot availablenot availablecondition_1
3D HNCAnot availablenot availablecondition_1
3D HN(CO)CAnot availablenot availablecondition_1
3D HNHAnot availablenot availablecondition_1
3D HNHBnot availablenot availablecondition_1
3D HNCOnot availablenot availablecondition_1
3D HNCACBnot availablenot availablecondition_1
3D CBCA(CO)NHnot availablenot availablecondition_1
3D HBHA(CO)NHnot availablenot availablecondition_1
3D 1H-15N HSQC-NOESYnot availablenot availablecondition_1
3D 1H-13C HSQC-NOESYnot availablenot availablecondition_1
3D 1H-13C NOESY-HMQCnot availablenot availablecondition_1
3D HCCH-TOCSYnot availablenot availablecondition_1
2D 1H-1H 13C-FILTERED NOESYnot availablenot availablecondition_1
2D 1H-1H 15N-FILTERED NOESYnot availablenot availablecondition_1

Software:

VNMR v6.1 - spectra collecting

NMRPipe v2.1 - spectra processing

Sparky v3.06 - spectra analysis, signal assignment

NMRTABLE v2.0 - analysis of resonance assignment

NMR spectrometers:

  • Varian UNITY 600 MHz
  • Varian INOVA 600 MHz
  • Varian INOVA 800 MHz

Related Database Links:

BMRB 17096 19563 19564 19565 19566 19567 7195
PDB
DBJ BAG35526 BAG56693 BAG59770 BAJ20267
EMBL CAA23765 CAA25409
GB AAA58484 AAA58485 AAH00192 AAH03584 AAH70280
PRF 0905195A 0906214A
REF NP_000782 NP_001277283 NP_001277286 XP_001099963 XP_001110551
SP P00374