Dihydrofolate+Reductase+(Plasmodium+Falciparum+3D7)

 Dihydrofolate Reducatase
 * Target (protein/gene name): **

 **NCBI Gene # or RefSeq#:**  Gene symbol: DHFR-TS  Gene ID: 9221804 http://www.ncbi.nlm.nih.gov/gene/9221804

 **Protein ID (NP or XP #) or Wolbachia#:**  PlasmoDB / PFD0830w  GeneDB / PFD0830w 155 http://tdrtargets.org/targets/view?gene_id=3944

 **Organism (including strain):**  Plasmodium Falciparum 3D7

 **Etiologic Risk Group (see link below):** Malaria caused by plasmodium parasites transmitted by mosquito bites Malaria affects some high risk groups more severely, including pregnant women, children under 5 years, people with HIV/AIDS and non-immune traveller Falciparum malaria is particularly frequent in sub-Saharan Africa and Oceania. http://www.who.int/malaria/high_risk_groups/en/

 ***Background/Disease Information:**  Plasmodium Falciparum is a protozoam parasite that causes malaria in humans. Also referred to as Falciparum Malaria, this form of Malaria represents the most severe form of malaria, with the highest rates of complications and morality http://www.niaid.nih.gov/topics/Malaria/understandingMalaria/Pages/default.aspx http://www.proteopedia.org/wiki/index.php/Malarial_Dihydrofolate_Reductase_as_Drug_Target

 **Essentiality of this protein:**  Yes, this protein has essentiality. It serves as probable target against Plasmodium Falciparum, which causes Malaria. http://www.ncbi.nlm.nih.gov/pubmed/15796007 http://tdrtargets.org/targets/view?gene_id=3944

 **Complex of proteins?:** Reductase(EC 1)

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Druggable Target:** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Druggability Index: 1 <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">247 chemical compunds associated with this gene. http://tdrtargets.org/targets/view?gene_id=3944

<span style="font-family: 'Arial Black',Gadget,sans-serif;">EC#: <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> 1.5.1.3

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Link to BRENDA EC# page:** <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Type of Enzyme Assay: Spectrophotometric http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/dihydrofolatereductase.pdf http://tdrtargets.org/targets/view?gene_id=3944

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **link to Sigma (or other company) page for assay or assay reagents (substrates)** <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[] <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **link (or citation) to paper that contains assay information** <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;">**List cost and quantity of substrate reagents and supplier** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> __Supplier:__ Sigma <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Potassium Phosphate Monobasic-(100g): $19.90 <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> B-Nicotinamide adenine dinucleotide 2'-phosphate reduced tetrasodium salt-(1mg): $148 <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Dihydrofolic Acid-(10mg): $71.60 <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Bovine Serum Albumin Solution-(10g): $73.60

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;">PDB # <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[|1J3I] <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]
 * <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Structure Available (PDB or Homology model) **

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Current Inhibitors:** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Dihydrofolate Reductase Inhibitors- <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Aminopterin <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Brodimoprim <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Iclaprim <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Methotrexate <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Premetrexed <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Proguanil <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Pralatrexate <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Proguanil <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Pyrimethamine <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Tetroxoprim <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Trimethoprim <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Trimethoprim/polumyxin <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Trimetrexate

http://www.scbt.com/chemicals-table-dhfr_inhibitors.html http://www.pharmacorama.com/en/Sections/DNA-RNA-biosynthesis-5.php

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Expression Information (has it been expressed in bacterial cells):** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">DHFR has been isolated and expressed in E.coli through the use of a plasmid(i.e pSV2-dhfr) <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[] http://www.jbc.org/content/268/29/21637.full.pdf+html

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Purification Method:** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> DHFR was purified by methotrexate-Sepharose affinity chromatography. <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> DHFR migrates as a single band on SDS-PAGE gel with a mass of -30 kDa <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Image of protein (PyMol with features delineated and shown separately):**



<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **DHFR crystal protein structure.(PDB:1J3I)** <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):** <span style="font-family: Arial,Helvetica,sans-serif;"> MMEQVCDVFDIYAICACCKVESKNEGKKNEVFNNYTFRGLGNKGVLPWKCNSLDMKYFCAVTTYVNESKY <span style="font-family: Arial,Helvetica,sans-serif;"> EKLKYKRCKYLNKETVDNVNDMPNSKKLQNVVVMGRTSWESIPKKFKPLSNRINVILSRTLKKEDFDEDV <span style="font-family: Arial,Helvetica,sans-serif;"> YIINKVEDLIVLLGKLNYYKCFIIGGSVVYQEFLEKKLIKKIYFTRINSTYECDVFFPEINENEYQIISV SDVYTSNNT <span style="font-family: Arial,Helvetica,sans-serif;"> TLDFIIYKKTNNKMLNEQNCIKGEEKNNDMPLKNDDKDTCHMKKLTEFYKNVDKYKINYEN DDDDEEEDDFVYF <span style="font-family: Arial,Helvetica,sans-serif;"> NFNKEKEEKNKNSIHPNDFQIYNSLKYKYHPEYQYLNIIYDIMMNGNKQSDRTGVGV LSKFGYIMKFDLSQYFPL <span style="font-family: Arial,Helvetica,sans-serif;"> LTTKKLFLRGIIEELLWFIRGETNGNTLLNKNVRIWEANGTREFLDNRKLFH REVNDLGPIYGFQWRHFGAEYTNMYDN <span style="font-family: Arial,Helvetica,sans-serif;"> YENKGVDQLKNIINLIKNDPTSRRILLCAWNVKDLDQMALPPC HILCQFYVFDGKLSCIMYQRSCDLGLGVPFNIASYSIF <span style="font-family: Arial,Helvetica,sans-serif;"> THMIAQVCNLQPAQFIHVLGNAHVYNNHIDSL KIQLNRIPYPFPTLKLNPDIKNIEDFTISDFTIQNYVHHEKISMDMAA <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> L**ength of your protein in Amino Acids:** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> 608 Amino Acids <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[]

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Molecular Weight of your protein in kiloDaltons using the** [|**Expasy ProtParam**]**<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> website ** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> 71737.2 http://web.expasy.org/cgi-bin/protparam/protparam

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> **Molar Extinction coefficient of your protein at 280 nm wavelength:** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Ext. coefficient 90620 Abs 0.1% (=1 g/l) 1.263, assuming all pairs of Cys residues form cystines <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> Ext. coefficient 89620 Abs 0.1% (=1 g/l) 1.249, assuming all Cys residues are reduced http://web.expasy.org/cgi-bin/protparam/protparam

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> ** TMpred graph Image ** ( <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">@http://www.ch.embnet.org/software/TMPRED_form.html <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">). Input your amino acid sequence to it.



<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> ***CDS Gene Sequence (paste as text only):** ATGATGGAACAAGTCTGCGACGTTTTCGATATTTATGCCATATGTGCATGTTGTAAGGTTGAAAGCAAAA ATGAGGGGAAAAAAAATGAGGTTTTTAATAACTACACATTTAGAGGTCTAGGAAATAAAGGAGTATTACC ATGGAAATGTAATTCCCTAGATATGAAATATTTTTGTGCAGTTACAACATATGTGAATGAATCAAAATAT GAAAAATTGAAATATAAGAGATGTAAATATTTAAACAAAGAAACTGTGGATAATGTAAATGATATGCCTA ATTCTAAAAAATTACAAAATGTTGTAGTTATGGGAAGAACAAGCTGGGAAAGCATTCCAAAAAAATTTAA ACCTTTAAGCAATAGGATAAATGTTATATTGTCTAGAACCTTAAAAAAAGAAGATTTTGATGAAGATGTT TATATCATTAACAAAGTTGAAGATCTAATAGTTTTACTTGGGAAATTAAATTACTATAAATGTTTTATTA TAGGAGGTTCCGTTGTTTATCAAGAATTTTTAGAAAAGAAATTAATAAAAAAAATATATTTTACTAGAAT AAATAGTACATATGAATGTGATGTATTTTTTCCAGAAATAAATGAAAATGAGTATCAAATTATTTCTGTT AGCGATGTATATACTAGTAACAATACAACATTGGATTTTATCATTTATAAGAAAACGAATAATAAAATGT TAAATGAACAAAATTGTATAAAAGGAGAAGAAAAAAATAATGATATGCCTTTAAAGAATGATGACAAAGA TACATGTCATATGAAAAAATTAACAGAATTTTACAAAAATGTAGACAAATATAAAATTAATTATGAAAAT GATGATGATGATGAAGAAGAAGATGATTTTGTTTATTTTAATTTTAATAAAGAAAAAGAAGAGAAAAATA AAAATTCTATACATCCAAATGATTTTCAAATATATAATAGCTTGAAATATAAATATCATCCTGAATACCA ATATTTAAATATTATTTATGATATTATGATGAATGGAAATAAACAAAGTGATCGAACGGGAGTAGGTGTT TTAAGTAAATTCGGATATATTATGAAATTTGATTTAAGTCAATATTTCCCATTATTAACTACGAAGAAAT TATTTTTAAGAGGAATTATTGAAGAATTGCTTTGGTTTATTAGAGGAGAAACAAATGGTAATACGTTGTT AAATAAGAATGTAAGGATATGGGAAGCTAATGGTACTAGGGAATTTTTAGATAATAGAAAATTATTTCAT AGAGAAGTTAACGATTTAGGACCTATTTATGGTTTTCAATGGAGACATTTCGGTGCTGAATATACAAATA TGTATGATAATTATGAAAATAAAGGAGTGGATCAATTAAAAAATATAATAAATTTAATTAAAAATGATCC TACAAGTAGAAGAATTCTTTTGTGTGCATGGAATGTAAAAGATCTTGACCAAATGGCATTACCTCCTTGT CATATTTTATGTCAGTTTTATGTTTTCGATGGGAAATTATCATGTATTATGTATCAAAGATCATGTGATT TAGGGCTAGGAGTACCTTTTAATATTGCTTCTTATTCTATTTTTACTCATATGATTGCACAAGTCTGTAA TTTGCAACCTGCGCAGTTCATACACGTTTTAGGAAATGCACATGTTTATAATAATCACATTGATAGTTTA AAAATTCAACTTAACAGAATACCCTATCCATTCCCAACACTTAAATTAAATCCAGATATTAAAAATATTG AAGATTTTACAATTTCGGATTTTACAATACAAAATTATGTTCATCATGAAAAAATTTCAATGGATATGGC TGCTTAA

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> ***GC% Content for gene:** 463/1827 GC%=27% http://www.genomicsplace.com/cgi-bin/gc_calculator.pl

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> ***CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;"> ***GC% Content for gene (codon optimized)** <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Primer for Lab PCR will be used.

<span style="font-family: 'Times New Roman','serif'; font-size: 16px;">**Internet Resources** <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[] <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[] <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[] <span style="color: #0000ff; font-family: 'Times New Roman','serif'; font-size: 16px;">[] http://web.expasy.org/cgi-bin/protparam/protparam http://tdrtargets.org/targets/view?gene_id=3944 http://www.jbc.org/content/257/22/13650.short http://www.addgene.org/vector-database/4256/ http://www.ncbi.nlm.nih.gov/gene/9221804 http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/dihydrofolatereductase.pdf