Target+-++dihydrofolate+reductase-thymidylate+synthase+(DHFR-TS)+(Leishmania+Major)

**Target:** Dihydrofolate Reductase-Thymidilate Synthase [Leishmania Major] **EC#:** 1.5.1.3 **NCBI Gene:** 159310 **Protein ID:** XP_001680857.1 **Organism:** Leishmania Major, Strain Friedlin **Etiologic Risk Group:** Risk Group 2 (Parasitic Agents) **Background/Disease Information:** Cutaneous Leishmaniasis Gene/Ortholog: eco2731 (OG4_10927); Phenotype: essential; Source study: gerdes Gene/Ortholog: Tb927.7.5480 (OG4_10927); Phenotype: significant gain of fitness in procyclic forms; Source study: alsford Gene/Ortholog: Tb927.7.5480 (OG4_10927); Phenotype: significant gain of fitness in differentiation of procyclic to bloodstream forms; Source study: alsford **BRENDA EC# Page:** http://tinyurl.com/DHFR-TS-BRENDA **NCBI Protein Page:** http://tinyurl.com/L-MAJ-DHFR-TS  - Sigma: http://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer/learning-center/assay-library/ec-number.html The DHFR assay mixture (1.0 mL) contained 25 pM Hzfolate, 0.1 mM NADPH, varying concentrations of inhibitors, 50 mM TES, 75 mM 2-mercaptoethanol, 1 mM EDTA, and 1.0 mg of BSA at pH m.0, 25 OC.” The reaction was initiated by addition of 0.5 unit of DHFR, and conversion of NADPH to NADP’ was monitored spectrophotometrically at 340 nm on a Cary-18 instrument. One unit of enzyme activity is defined as that amount of enzyme that produces 1 nmol of product/min. [3]
 * Essentiality: **
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Assay Information: **


 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Cost and Quantity of Substrate Reagents and Supplier: **


 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Homology Model: **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Closest PDB Entry:** 3INV <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Chain Used:** Chain A <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Query Coverage:** 98% <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Max Identities:** 348/523 (67%) <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Positives:** 403/523 (77%) <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Gaps:** 15/523 (2%)
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">- Pairwise Alignment: **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Second Closest PDB Entry:** 3IRM <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Chain Used:** Chain B <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Query Coverage:** 98% <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Max Identities:** 348/523 (67%) <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Positives:** 403/523 (77%) <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**- Gaps:** 15/523 (2%)
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">- Pairwise Alignment: **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**Current Inhibitors:** 22?? <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Weak activity with folate and no activity with pterins in Leishmania Major <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**Image of protein** (PyMol with features delineated and shown separately):
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Expression Information: **
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Purification Method: **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Courier New',Courier,monospace;"> 1 MSRAAARFKIPMPETKADFAFPSLRAFSIVVALDMQHGIGDGESIPWRVPEDMTFFKNQT <span style="font-family: 'Courier New',Courier,monospace;"> 61 TLLRNKKPPTEKKRNAVVMGRKTWESVPVKFRPLKGRLNIVLSSKATVEELLAPLPEGQR <span style="font-family: 'Courier New',Courier,monospace;">121 AAAAQDVVVVNGGLAEALRLLARPLYCSSIETAYCVGGAQVYADAMLSPCIEKLQEVYLT <span style="font-family: 'Courier New',Courier,monospace;">181 RIYATAPACTRFFPFPPENAATAWDLASSQGRRKSEAEGLEFEICKYVPRNHEERQYLEL <span style="font-family: 'Courier New',Courier,monospace;">241 IDRIMKTGIVKEDRTGVGTISLFGAQMRFSLRDNRLPLLTTKRVFWRGVCEELLWFLRGE <span style="font-family: 'Courier New',Courier,monospace;">301 TSAQLLADKDIHIWDGNGSREFLDSRGLTENKEMDLGPVYGFQWRHFGADYKGFEANYDG <span style="font-family: 'Courier New',Courier,monospace;">361 EGVDQIKLIVETIKTNPNDRRLLVTAWNPCALQKMALPPCHLLAQFYVNTDTSELSCMLY <span style="font-family: 'Courier New',Courier,monospace;">421 QRSCDMGLGVPFNIASYALLTILIAKATGLRPGELVHTLGDAHVYRNHVDALKAQLERVP <span style="font-family: 'Courier New',Courier,monospace;">481 HAFPTLIFKEERQYLEDYELTDMEVIDYVPHPAIKMEMAVX <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**Amino Acid Length of Protein**: 520 <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**Molecular Weight of Protein Using Expasy ProtParam:** 58688.6 kDa <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**Molar Extinction Coefficient at 280 nm Wavelength:** 69955 or 69330 <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Amino Acid Sequence: **
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">TMpred Graph: **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1 ATGTCCAGGGCAGCTGCGAGGTTTAAGATTCCGATGCCGGAGACGAAGGCAGACTTTGCT <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">61 TTCCCCTCCCTGCGCGCCTTCTCCATCGTCGTGGCCCTCGATATGCAGCACGGCATCGGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">121 GACGGCGAGTCGATCCCGTGGCGGGTGCCGGAGGACATGACGTTCTTCAAGAACCAGACG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">181 ACGCTGCTGCGCAACAAGAAGCCGCCGACGGAGAAGAAGCGCAACGCCGTCGTGATGGGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">241 CGCAAGACTTGGGAGAGCGTCCCGGTAAAGTTCCGACCACTCAAGGGACGGCTGAACATC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">301 GTGTTATCCTCGAAGGCCACCGTCGAGGAGCTTCTGGCGCCGCTGCCGGAGGGACAGCGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">361 GCGGCGGCGGCGCAGGATGTGGTGGTGGTGAACGGCGGTCTGGCCGAGGCGCTCCGCCTC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">421 CTCGCACGCCCGCTGTACTGCAGCTCCATCGAGACAGCGTATTGCGTCGGTGGTGCGCAG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">481 GTTTACGCGGACGCCATGCTGTCGCCGTGCATCGAGAAACTGCAGGAAGTGTACCTGACC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">541 CGCATCTACGCGACGGCGCCTGCGTGTACGCGCTTCTTTCCGTTTCCGCCCGAGAACGCG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">601 GCCACGGCGTGGGACCTGGCGTCGTCTCAGGGACGCCGCAAGAGCGAGGCGGAGGGCCTC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">661 GAGTTCGAGATCTGCAAGTACGTGCCGCGCAACCACGAGGAGCGGCAGTACCTTGAGCTG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">721 ATTGACCGCATCATGAAGACGGGGATCGTGAAGGAGGACCGCACCGGCGTGGGCACCATC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">781 AGCCTCTTCGGCGCCCAGATGCGCTTCTCCCTACGCGACAACCGCCTGCCGCTGCTGACG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">841 ACGAAGCGTGTCTTCTGGCGCGGCGTGTGCGAGGAGCTGCTGTGGTTCCTGCGCGGGGAG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">901 ACGAGTGCGCAGCTGCTGGCAGACAAGGACATTCACATCTGGGACGGCAACGGTTCGCGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">961 GAGTTTCTCGACAGCCGCGGCTTGACAGAGAATAAGGAGATGGACCTCGGCCCTGTCTAC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1021 GGCTTCCAGTGGCGCCACTTCGGGGCAGATTACAAGGGGTTTGAAGCGAACTACGACGGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1081 GAAGGGGTGGACCAGATCAAGCTCATCGTGGAGACCATCAAGACGAACCCGAACGACCGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1141 CGCCTCCTAGTCACTGCCTGGAACCCGTGCGCGCTGCAAAAGATGGCGCTGCCGCCGTGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1201 CACTTGCTTGCTCAGTTCTACGTGAACACAGACACGAGCGAGCTATCCTGCATGTTGTAC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1261 CAGCGCTCGTGTGACATGGGTCTTGGCGTCCCCTTCAACATTGCCTCCTACGCGCTGCTC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1321 ACCATCCTCATTGCCAAGGCGACGGGTCTGCGGCCTGGTGAGCTTGTGCACACCCTCGGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1381 GACGCCCACGTCTACCGCAACCACGTTGATGCCCTCAAGGCGCAGCTCGAGCGAGTCCCG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1441 CACGCGTTCCCGACCCTCATCTTCAAGGAGGAGCGGCAGTACCTCGAGGACTACGAGTTG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1501 ACGGACATGGAGGTGATCGACTACGTTCCACACCCGGCGATCAAGATGGAGATGGCCGTA <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1561 TAG <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**GC% Content:** 63.08%
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px; line-height: 1.5;">CDS Gene Sequence: **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;"> 1 ATGTCTCGTGCCGCAGCCCGCTTTAAGATCCCGATGCCTGAGACCAAAGCGGACTTCGCG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;"> 61 TTCCCGTCCCTGCGTGCTTTCTCTATTGTAGTAGCGCTGGACATGCAGCACGGTATTGGT <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">121 GACGGCGAGTCTATTCCGTGGCGTGTACCGGAGGATATGACCTTTTTCAAGAATCAGACC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">181 ACTCTGCTGCGCAACAAAAAACCGCCGACTGAGAAAAAACGCAATGCGGTTGTTATGGGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">241 CGCAAAACTTGGGAATCTGTCCCTGTTAAATTCCGTCCACTCAAAGGTCGTCTGAACATC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">301 GTTCTGTCTTCTAAAGCGACTGTTGAGGAACTGCTCGCACCTCTCCCAGAAGGCCAGCGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">361 GCAGCGGCAGCACAAGATGTTGTTGTGGTTAACGGTGGCCTGGCGGAAGCCCTCCGCCTC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">421 CTCGCGCGTCCGCTCTACTGTAGCTCTATCGAGACCGCCTACTGCGTAGGTGGTGCACAG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">481 GTTTACGCAGATGCCATGCTCTCCCCGTGCATCGAGAAGCTGCAGGAAGTATACCTGACT <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">541 CGCATTTACGCGACGGCTCCGGCGTGCACTCGTTTCTTCCCGTTCCCTCCGGAAAACGCG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">601 GCGACTGCGTGGGATCTCGCGTCCTCTCAAGGTCGCCGTAAATCTGAAGCGGAAGGCCTG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">661 GAATTTGAAATCTGCAAATACGTTCCGCGTAATCACGAAGAGCGTCAGTATCTCGAACTC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">721 ATCGACCGTATCATGAAAACTGGCATCGTGAAAGAAGACCGTACCGGTGTTGGCACCATC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">781 AGCCTGTTTGGTGCTCAGATGCGTTTCTCTCTGCGTGACAACCGTCTGCCACTGCTGACT <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">841 ACGAAACGTGTGTTCTGGCGTGGTGTTTGTGAGGAGCTGCTGTGGTTCCTCCGCGGTGAG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">901 ACGTCCGCGCAACTCCTGGCCGACAAAGACATTCACATCTGGGACGGCAATGGTTCCCGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">961 GAGTTCCTGGACTCTCGTGGCCTCACCGAGAACAAGGAGATGGACCTGGGTCCGGTGTAC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1021 GGCTTTCAGTGGCGTCACTTCGGTGCGGACTACAAAGGTTTCGAAGCGAACTACGATGGC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1081 GAAGGTGTTGACCAGATCAAACTGATCGTTGAAACCATCAAAACCAACCCTAACGACCGT <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1141 CGTCTCCTGGTAACCGCGTGGAACCCGTGCGCCCTGCAAAAGATGGCGCTCCCGCCGTGT <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1201 CATCTGCTGGCGCAATTCTACGTGAACACCGACACCTCTGAACTGTCTTGCATGCTGTAC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1261 CAGCGTTCTTGCGACATGGGCCTGGGCGTTCCTTTCAACATCGCGTCTTACGCGCTCCTG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1321 ACCATTCTCATCGCGAAAGCCACGGGCCTGCGTCCGGGCGAACTGGTCCATACCCTGGGT <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1381 GACGCGCATGTTTACCGCAACCACGTTGATGCGCTGAAGGCCCAGCTCGAACGTGTTCCA <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1441 CATGCATTCCCGACCCTGATTTTCAAAGAGGAACGTCAATACCTGGAGGACTATGAACTG <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1501 ACTGATATGGAAGTGATCGATTATGTACCACATCCGGCGATTAAGATGGAAATGGCTGTC <span style="font-family: 'Courier New',Courier,monospace; font-size: 14px;">1561 TAA <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;"> <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">**GC% Content:** 55.21%
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Codon Optimized CDS Gene Sequence: **


 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Virtual Screening Work **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">
 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Non-Prepped Control Library: **


 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Library Contains the Following Ligands : **<span style="font-family: 'Times New Roman',Times,serif; font-size: 14px; line-height: 1.5;">KKT, MGH, 1WL, AHM, IDK, ASP, DHF, P65, MMV, WRA, DQ1, 2CY, TMQ, MTX, C50, UMP, and 1CY

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 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px; line-height: 1.5;">Undergraduate Research Forum Poster: **


 * <span style="font-family: 'Times New Roman',Times,serif; font-size: 14px;">Rel <span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">evant Literature, References: **

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">1. Parasites†-†Leishmaniasis. http://www.cdc.gov/parasites/leishmaniasis/ (accessed October 12).

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">2. Andriantsoanirina, V.; Durand, R.; Pradines, B.; Baret, E.; Bouchier, C.; Ratsimbasoa, A.; MÈnard, D., In vitro susceptibility to pyrimethamine of DHFR I164L single mutant Plasmodium falciparum. Malar J 2011, 10, 283.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">3. Arrebola, R.; Olmo, A.; Reche, P.; Garvey, E. P.; Santi, D. V.; Ruiz-Perez, L. M.; Gonzalez-Pacanowska, D., Isolation and characterization of a mutant dihydrofolate reductase-thymidylate synthase from methotrexate-resistant Leishmania cells. J Biol Chem 1994, 269 (14), 10590-6.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">4. Artimo, P.; Jonnalagedda, M.; Arnold, K.; Baratin, D.; Csardi, G.; de Castro, E.; Duvaud, S.; Flegel, V.; Fortier, A.; Gasteiger, E.; Grosdidier, A.; Hernandez, C.; Ioannidis, V.; Kuznetsov, D.; Liechti, R.; Moretti, S.; Mostaguir, K.; Redaschi, N.; Rossier, G.; Xenarios, I.; Stockinger, H., ExPASy: SIB bioinformatics resource portal. Nucleic Acids Res 2012, 40 (Web Server issue), W597-603.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">5. Atreya, C. E.; Johnson, E. F.; Irwin, J. J.; Dow, A.; Massimine, K. M.; Coppens, I.; Stempliuk, V.; Beverley, S.; Joiner, K. A.; Shoichet, B. K.; Anderson, K. S., A molecular docking strategy identifies Eosin B as a non-active site inhibitor of protozoal bifunctional thymidylate synthase-dihydrofolate reductase. J Biol Chem 2003, 278 (16), 14092-100.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">6. Bernstein, F. C.; Koetzle, T. F.; Williams, G. J.; Meyer, E. F.; Brice, M. D.; Rodgers, J. R.; Kennard, O.; Shimanouchi, T.; Tasumi, M., The Protein Data Bank: a computer-based archival file for macromolecular structures. J Mol Biol 1977, 112 (3), 535-42.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">7. Beverley, S. M.; Ellenberger, T. E.; Cordingley, J. S., Primary structure of the gene encoding the bifunctional dihydrofolate reductase-thymidylate synthase of Leishmania major. Proc Natl Acad Sci U S A 1986, 83 (8), 2584-8.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">8. Booth, R. G.; Selassie, C. D.; Hansch, C.; Santi, D. V., Quantitative structure-activity relationship of triazine-antifolate inhibition of Leishmania dihydrofolate reductase and cell growth. J Med Chem 1987, 30 (7), 1218-24.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">9. Chitnumsub, P.; Yuvaniyama, J.; Chahomchuen, T.; Vilaivan, T.; Yuthavong, Y., Crystallization and preliminary crystallographic studies of dihydrofolate reductase-thymidylate synthase from Trypanosoma cruzi, the Chagas disease pathogen. Acta Crystallogr Sect F Struct Biol Cryst Commun 2009, 65 (Pt 11), 1175-8.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">10. Chitnumsub, P.; Yuvaniyama, J.; Vilaivan, T.; Vanichtanankul, J.; Kamchonwongpaisan, S.; Yuthavong, Y., Structural basis of antifolate inhibition of Trypanosoma cruzi Dihydrofolate Reductase-Thymidylate Synthase. 2011.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">11. Ferrari, S.; Morandi, F.; Motiejunas, D.; Nerini, E.; Henrich, S.; Luciani, R.; Venturelli, A.; Lazzari, S.; CalÚ, S.; Gupta, S.; Hannaert, V.; Michels, P. A.; Wade, R. C.; Costi, M. P., Virtual screening identification of nonfolate compounds, including a CNS drug, as antiparasitic agents inhibiting pteridine reductase. J Med Chem 2011, 54 (1), 211-21.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">12. Gilbert, I. H., Inhibitors of dihydrofolate reductase in Leishmania and trypanosomes. Biochim Biophys Acta 2002, 1587 (2-3), 249-57.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">13. Heaslet, H.; Harris, M.; Fahnoe, K.; Sarver, R.; Putz, H.; Chang, J.; Subramanyam, C.; Barreiro, G.; Miller, J. R., Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim. Proteins 2009, 76 (3), 706-17.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">14. Hillcoat, B. L.; Nixon, P. F.; Blakley, R. L., Effect of substrate decomposition on the spectrophotometric assay of dihydrofolate reductase. Anal Biochem 1967, 21 (2), 178-89.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">15. Jones, G.; Willett, P.; Glen, R. C.; Leach, A. R.; Taylor, R., Development and validation of a genetic algorithm for flexible docking. J Mol Biol 1997, 267 (3), 727-48.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">16. Khabnadideh, S.; Pez, D.; Musso, A.; Brun, R.; PÈrez, L. M.; Gonz·lez-Pacanowska, D.; Gilbert, I. H., Design, synthesis and evaluation of 2,4-diaminoquinazolines as inhibitors of trypanosomal and leishmanial dihydrofolate reductase. Bioorg Med Chem 2005, 13 (7), 2637-49.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">17. Knighton, D. R.; Kan, C. C.; Howland, E.; Janson, C. A.; Hostomska, Z.; Welsh, K. M.; Matthews, D. A., Structure of and kinetic channelling in bifunctional dihydrofolate reductase-thymidylate synthase. Nat Struct Biol 1994, 1 (3), 186-94.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">18. Lipinski, C. A.; Lombardo, F.; Dominy, B. W.; Feeney, P. J., Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 2001, 46 (1-3), 3-26.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">19. Meek, T. D.; Garvey, E. P.; Santi, D. V., Purification and characterization of the bifunctional thymidylate synthetase-dihydrofolate reductase from methotrexate-resistant Leishmania tropica. Biochemistry 1985, 24 (3), 678-86.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">20. Nare, B.; Hardy, L. W.; Beverley, S. M., The roles of pteridine reductase 1 and dihydrofolate reductase-thymidylate synthase in pteridine metabolism in the protozoan parasite Leishmania major. In J Biol Chem, 1997; Vol. 272, pp 13883-91.

<span style="font-family: 'Times New Roman',Times,serif; font-size: 15px;">21. Schomburg, I.; Chang, A.; Schomburg, D., BRENDA, enzyme data and metabolic information. Nucleic Acids Res 2002, 30 (1), 47-9.