Target (protein/gene name):
Dihydrofolate Reducatase

NCBI Gene # or RefSeq#:
Gene symbol: DHFR-TS
Gene ID: 9221804

Protein ID (NP or XP #) or Wolbachia#:
PlasmoDB / PFD0830w
GeneDB / PFD0830w 155

Organism (including strain):
Plasmodium Falciparum 3D7

Etiologic Risk Group (see link below):
*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


Essentiality of this protein:
Yes, this protein has essentiality. It serves as probable target against Plasmodium Falciparum, which causes Malaria.

Complex of proteins?:

Druggable Target:
Druggability Index: 1
247 chemical compunds associated with this gene.

EC#:
1.5.1.3

Link to BRENDA EC# page:
http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.5.1.3

Capture.PNG



Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):

Type of Enzyme Assay: Spectrophotometric

link to Sigma (or other company) page for assay or assay reagents (substrates)
http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/dihydrofolatereductase.pdf
http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/cs0340bul.Par.0001.File.tmp/cs0340bul.pdf

link (or citation) to paper that contains assay information
http://www.clinchem.org/content/45/2/223.full

-- List cost and quantity of substrate reagents and supplier

Supplier: Sigma
Potassium Phosphate Monobasic-(100g): $19.90
B-Nicotinamide adenine dinucleotide 2'-phosphate reduced tetrasodium salt-(1mg): $148
Dihydrofolic Acid-(10mg): $71.60
Bovine Serum Albumin Solution-(10g): $73.60

Structure Available (PDB or Homology model)
-- PDB #
1J3I
http://www.pdb.org/pdb/explore/explore.do?structureId=1J3I


Current Inhibitors:
Dihydrofolate Reductase Inhibitors-
Aminopterin
Brodimoprim
Iclaprim
Methotrexate
Premetrexed
Proguanil
Pralatrexate
Proguanil
Pyrimethamine
Tetroxoprim
Trimethoprim
Trimethoprim/polumyxin
Trimetrexate

http://tdrtargets.org/targets/view?gene_id=3944
http://en.wikipedia.org/wiki/Category:Dihydrofolate_reductase_inhibitors
http://www.ncbi.nlm.nih.gov/gene?cmd=Retrieve&dopt=full_report&list_uids=9221804
http://www.ncbi.nlm.nih.gov/protein/XP_001351479.1

Expression Information (has it been expressed in bacterial cells):
http://www.addgene.org/vector-database/4256/
DHFR has been isolated and expressed in E.coli through the use of a plasmid(i.e pSV2-dhfr)

Purification Method:
http://www.jbc.org/content/268/29/21637.full.pdf+html
DHFR was purified by methotrexate-Sepharose affinity chromatography.
DHFR migrates as a single band on SDS-PAGE gel with a mass of -30 kDa

Image of protein (PyMol with features delineated and shown separately):
1j3i_bio_r_500.jpg
DHFR crystal protein structure.(PDB:1J3I)
http://www.pdb.org/pdb/explore/explore.do?structureId=1J3I

*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):

MMEQVCDVFDIYAICACCKVESKNEGKKNEVFNNYTFRGLGNKGVLPWKCNSLDMKYFCAVTTYVNESKY
EKLKYKRCKYLNKETVDNVNDMPNSKKLQNVVVMGRTSWESIPKKFKPLSNRINVILSRTLKKEDFDEDV
YIINKVEDLIVLLGKLNYYKCFIIGGSVVYQEFLEKKLIKKIYFTRINSTYECDVFFPEINENEYQIISV SDVYTSNNT
TLDFIIYKKTNNKMLNEQNCIKGEEKNNDMPLKNDDKDTCHMKKLTEFYKNVDKYKINYEN DDDDEEEDDFVYF
NFNKEKEEKNKNSIHPNDFQIYNSLKYKYHPEYQYLNIIYDIMMNGNKQSDRTGVGV LSKFGYIMKFDLSQYFPL
LTTKKLFLRGIIEELLWFIRGETNGNTLLNKNVRIWEANGTREFLDNRKLFH REVNDLGPIYGFQWRHFGAEYTNMYDN
YENKGVDQLKNIINLIKNDPTSRRILLCAWNVKDLDQMALPPC HILCQFYVFDGKLSCIMYQRSCDLGLGVPFNIASYSIF
THMIAQVCNLQPAQFIHVLGNAHVYNNHIDSL KIQLNRIPYPFPTLKLNPDIKNIEDFTISDFTIQNYVHHEKISMDMAA

http://www.ncbi.nlm.nih.gov/protein/XP_001351479.1

*length of your protein in Amino Acids:
608 Amino Acids
http://www.ncbi.nlm.nih.gov/protein/XP_001351479.1

Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website
71737.2

Molar Extinction coefficient of your protein at 280 nm wavelength:
Ext. coefficient 90620 Abs 0.1% (=1 g/l) 1.263, assuming all pairs of Cys residues form cystines
Ext. coefficient 89620 Abs 0.1% (=1 g/l) 1.249, assuming all Cys residues are reduced

TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
external image .TMPRED.12176.5235.gif

*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
*GC% Content for gene:
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized)