TargetF15-+Inosine+5+Monophosphate+Dehydrogenase+(Bacillus+anthracis)

***Target (protein/gene name)**: Inosine 5 Monophosphate dehygrogenase ***NCBI Gene # or RefSeq#**: GuaB, 1083772 ***Protein ID (NP or XP #) or Wolbachia#**: GI: 349587863 ***Organism (including strain):** Bacillus anthracis str.Ames Anthrax derives from the gram-positive bacterium //Bacillus anthracis// with symptoms ranging from a rash to acute respiratory syndrome depending on the method of contraction. //Bacillus anthracis// is a bacterium which is capable of bioterrorist weaponization and is listed as a Category A pathogen by the US government. General public vaccination against //B.anthracis// is unrealistic given associated expenses, and current medications for the most deadly form of contraction--inhalation-- are medically intensive, requiring IV therapy and the assemblage of massive drug stockpiles. Given //B.anthracis//’ capacity for aerosol weaponization, the FDA has announced the need to create a prophylactic drug capable of being mass administered to individuals who are thought to have been exposed. By targeting //B.anthracis// Ionosine-5-monophosphate dehydrogenase (BaI5MD), an enzyme that is essential in //B.anthracis// DNA and RNA synthesis, our research aims to virtually screen large databases of drug-like molecules in order to identify potential inhibitors of BaI5MD. After cloning the BaI5MD gene and purifying BaI5MD protein, molecules which are predicted to bind with high affinity to BaI5MD can be selectively tested //in vitro// using spectrophotometric UV enzyme assays. Successful identification of novel BaI5MD inhibitors will then allow in-vivo testing of inhibition, followed by human cell toxicity tests. These developments would introduce the potential formulation of pharmaceutical therapies specifically targeted against //B.anthracis,// ideally eliminating it as a bioterrorist threat.
 * Etiologic Risk Group (see link below): ** Category A pathogen, US Dept of Health
 * */Disease Information (sort of like the Intro to your Mini Research Write up): **

**Link to TDR Targets page (if present):** N/A [|Link to Gene Database page (NCBI], EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **Essentiality of this protein:** Essential Reference: **Complex of proteins?**: Tetramer 0.8 in T.cruzi, 1 in P.falciparum ***EC#**: 1.1.1.205 [|Link to BRENDA EC# page:] <span style="font-family: Arial,Helvetica,sans-serif;"> <span style="font-family: Arial,Helvetica,sans-serif;">50 mM Tris-HCl (pH 8.0), 100 mM KCl, 3 mM EDTA, and 1 mM DTT; Reactions were initiated by the addition of IMPDH to a final concentration of 50 nM. The production of NADH was measured by monitoring the increase in absorbance at 340 nm (ε = 6.22 mM−1 cm−1) using a Shimadzu BioSpec-1601, Hitachi U-2000, or Cary 100 Bio spectrophotometer. <span style="font-family: Arial,Helvetica,sans-serif;">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3836674/#R7 __Ligand(s)__ determined from virtual screening. __NADH__ catalog #: N4505 supplier: Sigma-Aldrich Cost: $67.30 for 100MG <span style="font-family: Arial,Helvetica,sans-serif;">-- PDB #: 3TSB <span style="font-family: Arial,Helvetica,sans-serif;"> 3USB - docked with IMP
 * <span style="font-family: Arial,Helvetica,sans-serif;">Is it a monomer or multimer as biological unit? (make prediction at @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): **
 * <span style="font-family: Arial,Helvetica,sans-serif;">Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **
 * <span style="font-family: Arial,Helvetica,sans-serif;">Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * <span style="font-family: Arial,Helvetica,sans-serif;">-- link to Sigma (or other company) page for assay (see Sigma links below) **
 * <span style="font-family: Arial,Helvetica,sans-serif;">-- -or link (or citation) to paper that contains assay information: **
 * <span style="font-family: Arial,Helvetica,sans-serif;">-- links to assay reagents (substrates) pages. **
 * <span style="font-family: Arial,Helvetica,sans-serif;">--- List cost and quantity of substrate reagents, supplier, and catalog # **
 * <span style="font-family: Arial,Helvetica,sans-serif;">Structure (PDB or Homology model) **

<span style="font-family: Arial,Helvetica,sans-serif;">**Current Inhibitors**: mycophenolic acid, tiazofurin, ribavirin, mizoribine, rapamycin

<span style="font-family: Arial,Helvetica,sans-serif;">**Expression Information (has it been expressed in bacterial cells)**: yas... <span style="font-family: Arial,Helvetica,sans-serif;">**Purification Method**: Ni-NTA (has 6X His Tag) <span style="font-family: Arial,Helvetica,sans-serif;">***Amino Acid Sequence (paste as text only - not as screenshot or as 'code')**: <span style="font-family: Arial,Helvetica,sans-serif;">mhhhhhhssgvdlgtenlyfqsnamweskfvkegltfddvllvpaksdvlprevsvktvl seslqlniplisagmdtvteadmaiamarqgglgiihknmsieqqaeqvdkvkrsesgvi sdpffltpehqvydaehlmgkyrisgvpvvnnlderklvgiitnrdmrfiqdysikisdv mtkeqlitapvgttlseaekilqkykieklplvdnngvlqglitikdiekviefpnsakd kqgrllvgaavgvtadamtridalvkasvdaivldtahghsqgvidkvkevrakypslni iagnvataeatkalieaganvvkvgigpgsicttrvvagvgvpqltavydcatearkhgi pviadggikysgdmvkalaagahvvmlgsmfagvaespgeteiyqgrqfkvyrgmgsvga mekgskdryfqegnkklvpegiegrvpykgpladtvhqlvgglragmgycgaqdleflre naqfirmsgaglleshphhvqitkeapnysl <span style="font-family: Arial,Helvetica,sans-serif;">***length of your protein in Amino Acids:** 511aa <span style="font-family: Arial,Helvetica,sans-serif;">**Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website**: 55kDa <span style="font-family: Arial,Helvetica,sans-serif;">**Molar Extinction coefficient of your protein at 280 nm wavelength**: 26485 <span style="font-family: Arial,Helvetica,sans-serif;">
 * <span style="font-family: Arial,Helvetica,sans-serif;">Image of protein (PyMol with features delineated and shown separately): **
 * <span style="font-family: Arial,Helvetica,sans-serif;">TMpred graph Image (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. **
 * <span style="font-family: Arial,Helvetica,sans-serif;">*CDS Gene Sequence (paste as text only): **

<span style="font-family: Arial,Helvetica,sans-serif;">ATGTGGGAATCTAAATTCGTTAAAGAGGGCCTCACCTTTGACGACGTCCTGCTCGTCCCG <span style="font-family: Arial,Helvetica,sans-serif;"> 61 GCAAAAAGCGATGTACTCCCTCGTGAGGTTTCTGTTAAAACCGTTCTGTCCGAATCCCTC <span style="font-family: Arial,Helvetica,sans-serif;"> 121 CAGCTGAATATCCCGCTGATCTCTGCGGGTATGGATACGGTGACTGAAGCTGACATGGCG <span style="font-family: Arial,Helvetica,sans-serif;"> 181 ATTGCGATGGCTCGCCAAGGCGGCCTCGGCATTATTCACAAAAACATGTCTATCGAGCAG <span style="font-family: Arial,Helvetica,sans-serif;"> 241 CAAGCTGAACAGGTTGACAAGGTTAAACGCTCTGAATCTGGCGTCATCTCCGACCCGTTC <span style="font-family: Arial,Helvetica,sans-serif;"> 301 TTCCTGACCCCGGAGCATCAAGTGTACGACGCCGAACACCTGATGGGTAAATACCGCATC <span style="font-family: Arial,Helvetica,sans-serif;"> 361 TCTGGTGTCCCTGTCGTGAACAACCTCGACGAACGTAAGCTGGTAGGTATCATCACTAAC <span style="font-family: Arial,Helvetica,sans-serif;"> 421 CGTGACATGCGTTTTATTCAGGACTACTCTATCAAGATTAGCGACGTTATGACGAAGGAA <span style="font-family: Arial,Helvetica,sans-serif;"> 481 CAGCTGATTACTGCGCCTGTTGGTACTACCCTCTCCGAGGCAGAAAAAATCCTGCAAAAA <span style="font-family: Arial,Helvetica,sans-serif;"> 541 TACAAAATTGAGAAACTGCCGCTCGTTGATAATAACGGTGTTCTCCAGGGTCTCATCACG <span style="font-family: Arial,Helvetica,sans-serif;"> 601 ATTAAAGACATTGAAAAAGTTATCGAGTTCCCTAACTCTGCGAAGGACAAACAGGGTCGT <span style="font-family: Arial,Helvetica,sans-serif;"> 661 CTCCTGGTTGGCGCTGCAGTGGGTGTGACGGCTGACGCAATGACTCGTATCGACGCGCTG <span style="font-family: Arial,Helvetica,sans-serif;"> 721 GTTAAAGCGTCTGTTGATGCAATTGTTCTGGACACGGCCCACGGCCACTCCCAGGGCGTA <span style="font-family: Arial,Helvetica,sans-serif;"> 781 ATCGACAAAGTGAAGGAGGTTCGTGCGAAATACCCTTCTCTCAACATTATCGCGGGCAAC <span style="font-family: Arial,Helvetica,sans-serif;"> 841 GTAGCCACCGCTGAGGCTACTAAAGCGCTGATTGAGGCGGGTGCGAACGTTGTAAAAGTT <span style="font-family: Arial,Helvetica,sans-serif;"> 901 GGTATTGGTCCGGGTTCCATTTGCACGACTCGTGTAGTCGCAGGCGTTGGTGTGCCGCAA <span style="font-family: Arial,Helvetica,sans-serif;"> 961 CTGACTGCGGTTTACGACTGCGCGACCGAAGCCCGTAAACATGGCATCCCGGTAATCGCA <span style="font-family: Arial,Helvetica,sans-serif;">1021 GACGGCGGCATCAAATACTCTGGCGATATGGTAAAGGCACTGGCTGCTGGTGCACACGTA <span style="font-family: Arial,Helvetica,sans-serif;">1081 GTTATGCTCGGCTCTATGTTCGCCGGTGTTGCAGAGTCTCCGGGCGAAACTGAAATCTAC <span style="font-family: Arial,Helvetica,sans-serif;">1141 CAGGGCCGTCAATTCAAAGTATACCGTGGTATGGGTTCTGTGGGCGCTATGGAGAAAGGT <span style="font-family: Arial,Helvetica,sans-serif;">1201 TCCAAAGACCGTTACTTCCAGGAGGGTAACAAAAAACTCGTGCCGGAAGGTATTGAAGGT <span style="font-family: Arial,Helvetica,sans-serif;">1261 CGTGTACCGTACAAAGGTCCGCTCGCTGACACTGTACACCAGCTCGTCGGTGGTCTCCGT <span style="font-family: Arial,Helvetica,sans-serif;">1321 GCGGGCATGGGTTATTGTGGTGCGCAAGACCTCGAATTCCTCCGTGAGAATGCGCAGTTC <span style="font-family: Arial,Helvetica,sans-serif;">1381 ATCCGCATGAGCGGTGCTGGTCTGCTGGAATCTCACCCTCACCACGTGCAGATCACGAAA <span style="font-family: Arial,Helvetica,sans-serif;">1441 GAGGCTCCGAATTACAGCCTGTAA

<span style="font-family: Arial,Helvetica,sans-serif;">***GC% Content for gene:** 53.2%


 * <span style="font-family: Arial,Helvetica,sans-serif;">*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): **

<span style="font-family: Arial,Helvetica,sans-serif;">TACTTCCAATCCATGTGGGAATCTAAATTCGTTAAAGAGGGCCTCACCTTTGACGACGTCCTGCTCGTCCCG <span style="font-family: Arial,Helvetica,sans-serif;">GCAAAAAGCGATGTACTCCCTCGTGAGGTTTCTGTTAAAACCGTTCTGTCCGAATCCCTC <span style="font-family: Arial,Helvetica,sans-serif;">CAGCTGAATATCCCGCTGATCTCTGCGGGTATGGATACGGTGACTGAAGCTGACATGGCG <span style="font-family: Arial,Helvetica,sans-serif;">ATTGCGATGGCTCGCCAAGGCGGCCTCGGCATTATTCACAAAAACATGTCTATCGAGCAG <span style="font-family: Arial,Helvetica,sans-serif;">CAAGCTGAACAGGTTGACAAGGTTAAACGCTCTGAATCTGGCGTCATCTCCGACCCGTTC <span style="font-family: Arial,Helvetica,sans-serif;">TTCCTGACCCCGGAGCATCAAGTGTACGACGCCGAACACCTGATGGGTAAATACCGCATC <span style="font-family: Arial,Helvetica,sans-serif;">TCTGGTGTCCCTGTCGTGAACAACCTCGACGAACGTAAGCTGGTAGGTATCATCACTAAC <span style="font-family: Arial,Helvetica,sans-serif;">CGTGACATGCGTTTTATTCAGGACTACTCTATCAAGATTAGCGACGTTATGACGAAGGAA <span style="font-family: Arial,Helvetica,sans-serif;">CAGCTGATTACTGCGCCTGTTGGTACTACCCTCTCCGAGGCAGAAAAAATCCTGCAAAAA <span style="font-family: Arial,Helvetica,sans-serif;">TACAAAATTGAGAAACTGCCGCTCGTTGATAATAACGGTGTTCTCCAGGGTCTCATCACG <span style="font-family: Arial,Helvetica,sans-serif;">ATTAAAGACATTGAAAAAGTTATCGAGTTCCCTAACTCTGCGAAGGACAAACAGGGTCGT <span style="font-family: Arial,Helvetica,sans-serif;">CTCCTGGTTGGCGCTGCAGTGGGTGTGACGGCTGACGCAATGACTCGTATCGACGCGCTG <span style="font-family: Arial,Helvetica,sans-serif;">GTTAAAGCGTCTGTTGATGCAATTGTTCTGGACACGGCCCACGGCCACTCCCAGGGCGTA <span style="font-family: Arial,Helvetica,sans-serif;">ATCGACAAAGTGAAGGAGGTTCGTGCGAAATACCCTTCTCTCAACATTATCGCGGGCAAC <span style="font-family: Arial,Helvetica,sans-serif;">GTAGCCACCGCTGAGGCTACTAAAGCGCTGATTGAGGCGGGTGCGAACGTTGTAAAAGTT <span style="font-family: Arial,Helvetica,sans-serif;">GGTATTGGTCCGGGTTCCATTTGCACGACTCGTGTAGTCGCAGGCGTTGGTGTGCCGCAA <span style="font-family: Arial,Helvetica,sans-serif;">CTGACTGCGGTTTACGACTGCGCGACCGAAGCCCGTAAACATGGCATCCCGGTAATCGCA <span style="font-family: Arial,Helvetica,sans-serif;">GACGGCGGCATCAAATACTCTGGCGATATGGTAAAGGCACTGGCTGCTGGTGCACACGTA <span style="font-family: Arial,Helvetica,sans-serif;">GTTATGCTCGGCTCTATGTTCGCCGGTGTTGCAGAGTCTCCGGGCGAAACTGAAATCTAC <span style="font-family: Arial,Helvetica,sans-serif;">CAGGGCCGTCAATTCAAAGTATACCGTGGTATGGGTTCTGTGGGCGCTATGGAGAAAGGT <span style="font-family: Arial,Helvetica,sans-serif;">TCCAAAGACCGTTACTTCCAGGAGGGTAACAAAAAACTCGTGCCGGAAGGTATTGAAGGT <span style="font-family: Arial,Helvetica,sans-serif;">CGTGTACCGTACAAAGGTCCGCTCGCTGACACTGTACACCAGCTCGTCGGTGGTCTCCGT <span style="font-family: Arial,Helvetica,sans-serif;">GCGGGCATGGGTTATTGTGGTGCGCAAGACCTCGAATTCCTCCGTGAGAATGCGCAGTTC <span style="font-family: Arial,Helvetica,sans-serif;">ATCCGCATGAGCGGTGCTGGTCTGCTGGAATCTCACCCTCACCACGTGCAGATCACGAAA <span style="font-family: Arial,Helvetica,sans-serif;">GAGGCTCCGAATTACAGCCTGTAACAGTAAAGGTGGATA


 * <span style="font-family: Arial,Helvetica,sans-serif;">*GC% Content for gene (codon optimized): 52.1% **

<span style="font-family: Arial,Helvetica,sans-serif;">[|link to DNA Works output][|text file]
 * <span style="font-family: Arial,Helvetica,sans-serif;">Do Not Need this info for Spring (but still copy these lines to your Target page for now) **
 * <span style="font-family: Arial,Helvetica,sans-serif;">Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **

<span style="font-family: Arial,Helvetica,sans-serif;">
 * <span style="font-family: Arial,Helvetica,sans-serif;">Primer design results for 'tail' primers (this is just 2 sequences): **