Target-Histidine+Phosphatase+(Francisella+Tularensis)

**Target** : //__Histindine Acid Phosphotase__// **EC #:** __//3.1.3.2//__ **Length of Protein:**//__342 residues__// **Molecular Weight:**
 * Organism** :// __Francisella Tularensis subsp. holarctica LVS__ //
 * NCBI GENE # or RefSeq#: ** //__5523568__//

IC50 determination of top compounds DSF analysis of ligands (machine in Robertus lab) pharmacophore searching in ICM http://www.molsoft.com/gui/cheminformatics-tutorials.html#tutorial-3D-pharmacophore Ensemble docking in ICM and GOLD -- i.e. docking against multiple receptor conformations. How many PDB structures are there for your enzyme?
 * Possible Items to Do for this project: -- Dr. B**

//In silico// alteration of one amino acid residue that is most likely to be mutated in a resistant form of the bacterium and then virtual screening against that structure with your same top hits -- this mutant version could then be grown up and test in wet lab against the same compounds Molecular dynamics simulations and re-scoring of top ligands crystallization

__Links__: [|Original Article] [|PDB Article]
 * Crystallization Info: **

__Temperature__: 293K __Final Optimized condition__: 10%(v/v) //Tascimate//, 0.1 M HEPES pH 7.0 and 19%(w/v) PEG 3350 __Extra Info__: Equal volumes of the protein (1.5 ml) and the reservoir (1.5 ml) were mixed and allowed to equilibrate with 1.0 ml of reservoir for 24–48 h. __**NOTE**__: //Tacsimate// (Hampton Research, HR2-755) is a mixture of organic acids that includes sodium malonate, sodium acetate, triammonium citrate, succinic acid, dl-malic acid and sodium formate
 * WITH L(+)-tartrate ion**

These crystals were grown in sitting drops at room temperature using a protein stock solution of 10 mg/mL enzyme in 50 mM sodium acetate pH 6.0, and reservoir solutions consisting of 0.05 – 0.2 M Bis-Tris buffer pH 5.0, and 1.6 – 2.0 M ammonium sulfate
 * Inhibitor Free**:

Large tetragonal crystals were grown in sitting drops at room temperature using reservoir solutions of 0.05– 0.20 M Bis-Tris pH 5.0 – 6.5 and 17 – 25 % PEG 3350. The Pi complex was formed by soaking a crystal for 30 minutes in 21 % (w/v) PEG 3350, 0.1 M Bis-Tris pH 6.0, 25% (v/v) PEG 200, and 20 mM Pi (from a stock solution of 100 mM KH2PO4 adjusted to pH 6.0). The crystal was then picked up with a Hampton loop and plunged into liquid nitrogen.
 * With Phosphate ION**

__ TACTTCCAATCCATGAAGAAAATCTTTGTTAG __ ** Reverse Primer: ** __ TATCCACCTTTACTG TTATTTCTGGAATTTTTCG __
 * Forward Primer: **

**Background/Disease:** **Image:**

>gi|89255449:32306-33361 Francisella tularensis subsp. holarctica LVS chromosome, complete genome ATGAAAAAAATTTTTGTGTCATTTACATTATTGTTTTTTTTGATACCTGTAGGCTACTCAAGTAAGCTGATTT TTGTATCGATGATAACACGTCATGGTGATAGAGCACCTTTTGCAAATATCGAAAATGCTAATTATAGCTGGG GAACTGAGCTATCTGAGCTTACCCCTATTGGGATGAATCAAGAATATAATTTAGGACTTCAGCTAAGAAAAA GATATATAGATAAATTTGGTCTTTTACCAGAGCATTATGTTGATCAAAGTATCTATGTGCTATCAAGTCACACA AATCGTACAGTGGTTAGTGCTCAAAGTTTGCTTATGGGTCTATATCCAGCTGGTACAGGACCGCTAATTGG TGATGGTGATCCAGCAATTAAAGATAGATTTCAACCAATACCAATAATGACCTTGTCAGCAGATTCTCGACT TATACAGTTTCCTTATGAACAATATTTAGCTGTATTAAAAAAATATGTCTATAACTCACCAGAGTGGCAAAACA AGACAAAAGAAGCTGCACCAAATTTTGCAAAATGGCAGCAAATACTAGGTAATAGAATCTCAGGATTAAATGA TGTTATTACAGTAGGAGATGTTTTGATTGTTGCTCAAGCACATGGTAAACCTTTACCAAAGGGTTTATCTCAA GAAGATGCTGATCAGATAATTGCTCTTACAGATTGGGGACTAGCGCAGCAGTTTAAATCACAAAAAGTATCC TATATCATGGGTGGTAAATTGACTAATAGAATGATTGAAGATCTAAATAATGCCGTTAATGGTAAATCTAAGTAT AAAATGACTTACTATTCAGGGCATGATCTGACACTTTTAGAAGTTATGGGTACTTTAGGCGTACCACTAGATA CAGCGCCAGGATATGCAAGTAATCTAGAAATGGAACTTTATAAAGATGGCGATATCTATACGGTTAAGCTTAG ATACAATGGTAAGTATGTCAAACTACCTATAATGGATAAAAATAATAGTTGCAGTTTAGATGCTCTAAATAAGTAT <span style="background-color: transparent; color: #000000; font-family: 'Courier New',Courier,monospace; font-size: 14px; text-decoration: none; vertical-align: baseline;">ATGCAGAGTATTAATGAGAAGTTTCAAAAATAA
 * CDS GENE SEQUENCE: **

<span style="background-color: transparent; color: #000000; font-family: Arial; font-size: 14.6667px; text-decoration: none; vertical-align: baseline;">>gi|89255478|ref|YP_512839.1| acid phosphatase [Francisella tularensis subsp. holarctica LVS] <span style="background-color: transparent; color: #000000; font-family: Courier New; font-size: 14.6667px; text-decoration: none; vertical-align: baseline;">MKKIFVSFTLLFFLIPVGYSSKLIFVSMITRHGDRAPFANIENANYSWGTELSELTPIGMNQEYNLGLQLRKRYIDKFGLLPEHYVDQSIYVLSSHTNRTVVSAQSLLMGLYPAGTGPLIGDGDPAIKDRFQPIPIMTLSADSRLIQFPYEQYLAVLKKYVYNSPEWQNKTKEAAPNFAKWQQILGNRISGLNDVITVGDVLIVAQAHGKPLPKGLSQEDADQIIALTDWGLAQQFKSQKVSYIMGGKLTNRMIEDLNNAVNGKSKYKMTYYSGHDLTLLEVMGTLGVPLDTAPGYASNLEMELYKDGDIYTVKLRYNGKYVKLPIMDKNNSCSLDALNKYMQSINEKFQK
 * AMINO ACID SEQUENCE: **

<span style="background-color: transparent; color: #000000; font-family: Courier New; font-size: 14.6667px; text-decoration: none; vertical-align: baseline;">NIAID Video

@http://www.youtube.com/watch?v=itZE_y5sCFc&feature=youtu.be