Glutathione+reductase+Plasmodium+falciparum+3D7

=//**Plasmodium falciparum 3D7-**// **Glutathione Reductase**=

Plasmodium falciparum is the most deadly of the five Plasmodium species that cause human malaria. Malaria has a massive impact on human health; it is the worlds second biggest killer. Around 300 million clinical cases occur each year resulting in between 1.5 - 2.7 million deaths annually, with the majority of cases being present in sub- saharan Africa. It is estimated that 3,000 children under the age of five years fall victim to malaria each day. Additonally, around 40% of the worlds population are at risk. Is it a monomer or multimer as biological unit ** ? (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html):
 * Target (protein/gene name): Plasmodium falciparum 3D7- Glutathione Reductase
 * NCBI Gene # or RefSeq#:811773
 * *Protein ID (NP or XP #) or Wolbachia#: ** Accession CZT99905.1
 * *Organism (including strain): ** Plasmodium falciparum 3D7 (isolate 3D7)
 * Etiologic Risk Group (see link below): RG2 **
 * */ Disease Information (sort of like the Intro to your Mini Research Write up): **
 * Link to TDR Targets page (if present): ** [|TDR Targets page]
 * Link to Gene Database page: **[|NCBI]
 * Essentiality of this protein: ** Essential for glutathione metabolism and reductase pathway
 * Complex of proteins?: ** No, Dimer formed from two identical monomers
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): ** [|Papers for druggable target]
 * *EC#: ** 1.8.1.7
 * Link to BRENDA EC# page: **http://www.brenda-enzymes.org/enzyme.php?ecno=1.8.1.7

http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/glutathionereductase.pdf http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/glutathionereductase.pdf 100 mM Potassium Phosphate Buffer with 3.4 mM Ethylenediaminetetraacetic Acid 30 mM Glutathione Substrate Solution (GSSG) 0.8 mM ß-Nicotinamide Adenine Dinucleotide Phosphate, Reduced Form Solution 1.0% (w/v) Bovine Serum Albumin Glutathione Reductase Enzyme Solution -- PDB # or closest PDB entry if using homology model: 3SQP for human and 1ONF for plasmodium (1R(S),2R(S),3S(R),4S(R))-2,3-dihydroxycyclo-hexane-1,4-diyl dinitrate mutant enzyme glutathione reductase-trypanothine reductase, expression in Escherichia coli [|(Brenda)] ammonium sulfate fractionation, 2',5'-ADP Sepharose 4B affinity chromatography and Sephadex G-200 gel filtration [|(Brenda)]
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * -- link to Sigma (or other company ) page for assay (see Sigma links below) **
 * -- -or link (or citation) to paper that contains assay information **
 * -- links to assay reagents (substrates) pages. **
 * -- List cost and quantity of substrate reagents, supplier, and catalog # **
 * Structure (PDB or Homology model **
 * Current Inhibitors: **
 * Expression Information (has it been expressed in bacterial cells):**
 * Purification Method:**
 * Image of protein (PyMol with features delineated and shown separately): **



MVYDLIVIGGGSGGMAAARRAARHNAKVALVEKSRLGGTCVNVGCVPKKIMFNAASVHDI LENSRHYGFDTKFSFNLPLLVERRDKYIQRLNNIYRQNLSKDKVDLYEGTASFLSENRIL IKGTKDNNNKDNGPLNEEILEGRNILIAVGNKPVFPPVKGIENTISSDEFFNIKESKKIG IVGSGYIAVELINVIKRLGIDSYIFARGNRILRKFDESVINVLENDMKKNNINIVTFADV VEIKKVSDKNLSIHLSDGRIYEHFDHVIYCVGRSPDTENLKLEKLNVETNNNYIVVDENQ RTSVNNIYAVGDCCMVKKSKEIEDLNLLKLYNEERYLNKKENVTEDIFYNVQLTPVAINA GRLLADRLFLKKTRKTNYKLIPTVIFSHPPIGTIGLSEEAAIQIYGKENVKIYESKFTNL FFSVYDIEPELKEKTYLKLVCVGKDELIKGLHIIGLNADEIVQGFAVALKMNATKKDFDE TIPIHPTAAEEFLTLQPWMKX
 * Fig 1.** Protein 1ONF PyMol image
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **


 * *length of your protein in Amino Acids: ** 500 aa
 * Molecular Weight of your protein in kiloDaltons: ** 56.492 kDa
 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 99115M^-1 cm^-1
 * TMpred graph Image **


 * Fig 2. ** TMpred graph of the chains in glutathione reductase protein of Plasmodium Falciparum.

ATGGTTTACGACCTCATCGTTATCGGTGGTGGTTCTGGCGGTATGGCTGCGGCACGTCGT GCGGCTCGTCACAACGCGAAAGTTGCTCTGGTTGAAAAATCTCGTCTGGGCGGCACCTGC GTAAACGTAGGTTGCGTTCCGAAGAAGATCATGTTCAACGCGGCTAGCGTTCACGATATC CTGGAGAATTCCCGTCATTACGGTTTCGATACCAAATTCAGCTTTAACCTGCCGCTGCTC GTTGAGCGTCGTGACAAATACATCCAGCGTCTGAACAACATTTATCGTCAAAACCTCTCC AAAGACAAGGTTGACCTGTACGAGGGTACTGCGTCTTTCCTCTCCGAAAACCGCATTCTC ATCAAGGGTACGAAAGATAATAACAACAAGGACAACGGTCCGCTGAATGAGGAAATTCTG GAGGGCCGTAACATCCTGATCGCAGTAGGCAACAAGCCAGTTTTCCCGCCTGTGAAAGGT ATTGAGAACACCATCTCTTCTGATGAGTTCTTCAACATCAAAGAATCCAAAAAGATTGGC ATCGTTGGCAGCGGTTACATCGCGGTTGAGCTGATCAACGTGATCAAACGCCTCGGCATT GACTCTTACATTTTTGCGCGTGGCAACCGTATCCTGCGCAAATTCGACGAATCTGTGATT AATGTGCTCGAGAATGACATGAAAAAGAACAATATCAACATTGTGACCTTCGCGGACGTT GTTGAAATCAAGAAAGTTTCTGACAAGAACCTGAGCATTCACCTCAGCGACGGTCGTATC TACGAGCACTTCGACCACGTCATCTATTGCGTAGGTCGTTCCCCGGACACCGAAAATCTG AAACTGGAAAAACTGAACGTTGAGACCAACAATAACTACATTGTTGTCGACGAGAACCAG CGCACCTCTGTAAATAACATCTATGCAGTCGGTGACTGCTGCATGGTTAAGAAGTCTAAA GAGATCGAAGACCTGAACCTCCTGAAGCTGTATAACGAGGAGCGTTACCTCAACAAAAAG GAAAATGTTACCGAAGACATCTTCTATAACGTCCAGCTGACCCCGGTTGCTATCAATGCT GGTCGCCTCCTCGCGGATCGTCTGTTCCTCAAGAAGACCCGTAAAACCAACTACAAACTG ATCCCGACCGTTATTTTCTCCCACCCGCCGATTGGTACGATCGGTCTGTCCGAGGAAGCG GCGATCCAGATCTACGGTAAGGAGAACGTTAAGATCTATGAAAGCAAATTTACGAACCTG TTTTTCTCTGTTTATGACATCGAACCGGAACTGAAAGAAAAAACGTATCTCAAACTCGTG TGTGTTGGTAAAGACGAACTGATTAAAGGCCTGCACATCATCGGCCTGAATGCAGACGAA ATCGTTCAGGGTTTCGCCGTAGCGCTCAAAATGAACGCGACCAAGAAGGACTTCGATGAA ACCATCCCAATCCACCCAACTGCGGCGGAAGAGTTTCTCACTCTGCAGCCGTGGATGAAG TGA
 * *CDS Gene Sequence (paste as text only): **
 * *GC% Content for gene (codon optimized): ** unable to to identify information


 * Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **
 * ( link to DNA Works output text file - ** that should be saved in your Google Docs folder after you did the primer design protocol)


 * Primer design results for 'tail' primers (this is just 2 sequences): **

Tail TACTTCCAATCC ATG (3-5)

Start codon: (ATG)

Start bps GTTTACGATCTCATCGTTATCGGTGGC

3-5 BEGINNING

TACTTCCAATCC ATG GTTTACGATCTCATCGTTATCGGTGGC (27)

End bps CTGACTCTGCAACCGTGGATGAAG

Stop codon:TTA

Tails TATCCACCTTTACTG (3-5)

5-3 DIRECTION REVERSE COMPLEMENT

TATCCACCTTTACTG TTA CTTCATCCACGGTTGCAGAGTCAG (24)

Tail TACTTCCAATCC ATG (3-5) Start codon: (ATG) Start bps GTTTACGATCTCATCGTTATCGGTGGC 3-5 BEGINNING TACTTCCAATCC __ATG__ GTTTACGATCTCATCGTTATCGGTGGC (27)

End bps CTGACTCTGCAACCGTGGATGAAG Stop codon:TTA Tails TATCCACCTTTACTG (3-5) 5-3 DIRECTION REVERSE COMPLEMENT TATCCACCTTTACTG __TTA__ CTTCATCCACGGTTGCAGAGTCAG (24)

0.4Um Oligo conc, 50Mm Na+, 0.3Mm Dntp forward with 27 start bp; 42 length; GC 45.2% 0mM: 65.4; 1.5mM: 73; 2mM 73.3; 4mM: 74mM; 6mM: 74.4

reverse with 24 ending bp; 42 length; GC 45.2% 0mM: 65.2; 1.5mM: 72.9; 2mM 73.2; 4mM: 73.9mM; 6mM: 74.2