Target+-+protein-tyrosine-phosphatase+(Toxoplasma+gondii)


 * Target (protein/gene name):** dual specificity phosphatase, catalytic domain-containing protein


 * NCBI Gene # or RefSeq#:** 7895273


 * Protein ID (NP or XP #) or Wolbachia#:** EPT31998.1


 * Organism (including strain):** Toxoplasma gondii (ME49 )


 * Etiologic Risk Group (see link below):** Risk Group 2 Parasitic Agent

Toxoplasmosis is a disease found throughout the word that is caused by a single-celled, obligate intracellular, parasitic parasitic protozoan, T. gondii. The way people get the disease is from ingestion of any under-cooked, contaminated meat, and not washing hands thoroughly after handling it. Symptoms of toxoplasmosis including swollen lymph glands that feel like the flu, or long lasting pains and muscle aches.
 * Background/ Disease Information (sort of like the Intro to your Mini Research Write up):**


 * Link to TDR Targets page (if present):** http://tdrtargets.org/targets/view?gene_id=259586

http://www.ncbi.nlm.nih.gov/gene/?term=TGME49_021590
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)**

"This entry represents dual specificity protein-tyrosine phosphatases. Ser/Thr and Tyr dual specificity phosphatases are a group of enzymes with both Ser/Thr (EC:3.1.3.16) and tyrosine specific protein phosphatase (EC:3.1.3.48) activity able to remove both the serine/threonine or tyrosine-bound phosphate group from a wide range of phosphoproteins, including a number of enzymes which have been phosphorylated under the action of a kinase. Dual specificity protein phosphatases (DSPs) regulate mitogenic signal transduction and control the cell cycle. The crystal structure of a human DSP, vaccinia H1-related phosphatase (or VHR), has been determined at 2.1 angstrom resolution [PMID: 8650541]. A shallow active site pocket in VHR allows for the hydrolysis of phosphorylated serine, threonine, or tyrosine protein residues, whereas the deeper active site of protein tyrosine phosphatases (PTPs) restricts substrate specificity to only phosphotyrosine. Positively charged crevices near the active site may explain the enzyme's preference for substrates with two phosphorylated residues. The VHR structure defines a conserved structural scaffold for both DSPs and PTPs. A "recognition region" connecting helix alpha1 to strand beta1, may determine differences in substrate specificity between VHR, the PTPs, and other DSPs." Source: Dual specificity phosphatase, catalytic domain (IPR000340) (http://www.ebi.ac.uk/interpro/entry/IPR000340)
 * Essentiality of this protein:**


 * Complex of proteins?:** secrete phosphotyrosine phosphatase

Cdc25 Source: Protein tyrosine phosphatases: mechanisms of catalysis and regulation (http://www.sciencedirect.com/science/article/pii/S1367593198800951)
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):**


 * *EC#: **3.1.3.48

http://www.brenda-enzymes.org/php/result_flat.php4?ecno=3.1.3.48
 * Link to BRENDA EC# page:**


 * --** Show screenshot of BRENDA enzyme mechanism schematic

http://www.sigmaaldrich.com/catalog/search?interface=All&term=toxoplasma%20gondii%20phosphatase&N=0&focus=product&lang=en®ion=US
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):**
 * -- link to Sigma (or other company ) page for assay (see Sigma links below)**

http://www.sigmaaldrich.com/catalog/papers/10779791
 * -- -or link (or citation) to paper that contains assay information**

http://www.sigmaaldrich.com/catalog/product/sigma/s7821?lang=en®ion=US
 * -- links to assay reagents (substrates) pages.**

Substrate reagent: sulfadoxin [S7821] 10G: $68.50 25G: 169.00 Supplier: Sigma-Aldrich CAS Number: 2447-57-6
 * --- List cost and quantity of substrate reagents, supplier, and catalog #**

-- PDB # or closest PDB entry if using homology model: 2ISN (not very similar actually - Dr. B 091114) -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB Query Coverage: Max % Identities: % Positives Chain used for homology:
 * Structure Available (PDB or Homology model)**

two heat-stable proteins known as Inhibitor-1 (I-1) and Inhibitor-2 (I-2) Source: Protein Phosphatase Inhibitors ( http://www.emdmillipore.com/life-science-research/protein-phosphatase-inhibitors/c_Sc2b.s1L.woAAAEWyGEfVhTm)
 * Current Inhibitors:**

Inhibitory phosphorylation on Thr and Tyr located in the glycine-rich ATP anchor motif of cdks results in inactivation Source: Small molecule inhibitors of dual specificity protein phosphatases (http://www.nature.com/onc/journal/v19/n56/full/1204084a.html)

"The conditional expression system to generate a transgenic parasite line in which the levels of T. gondii AMA1 (TgAMA1) expression can be experimentally manipulated." Source: Conditional Expression of Toxoplasma gondii Apical Membrane Antigen-1 (TgAMA1) Demonstrates That TgAMA1 Plays a Critical Role in Host Cell Invasion ( http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1196342/)
 * Expression Information (has it been expressed in bacterial cells):**

"Four methods to separate and purify Toxoplasma gondii tachyzoites from in vivo and in vitro culture systems, including trypsin digestion, purification with a 3-μm filter, CF-11 cellulose purification, and Percoll purification." Source: Separation and purification of Toxoplasma gondii tachyzoites from in vitro and in vivo culture systems (http://www.ncbi.nlm.nih.gov/pubmed/22033076)
 * Purification Method :**


 * Image of protein (PyMol with features delineated and shown separately):**

">2ISN:A|PDBID|CHAIN|SEQUENCE SLKKVITVNEWYTTTVAATMLGRRPTDEDAILVSAPATSRPNVRIKAVFDGHAGEATSQYCAKHAAKHLGKLSEFTFAEV KKACLSLDAEIIRKLGPKHVAGSTGIIVAIERLSAPVVENVVGREIVPRAHEETFVPLEKLIQEEEEAEHPELVGRYPRV PDVQQKTIPAGSFLVTAINIGDSRATLIHSDGGLTRLSKDHKPNHPTEASRIEKAGGSVETFDVPRVDGVLALSRAFGDS DFKMNPNLPPEEQKVIAVPDVRQFYALSSDLLLLACDGVYEPSGMDWAYVRDLTVAEMQRSKGDLEEVAARVMDYAYDMN SQDNISVMLVAFHNQEVEHPTAVYKVVSGQVDKVEWDVAGKKGN
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**


 * length of your protein in Amino Acids: 364**

39850.2 g/mol
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website:**

Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water. Ext. coefficient: 30035 M-1 cm-1 Abs 0.1% (=1 g/l) 0.754, assuming all pairs of Cys residues form cystines
 * Molar Extinction coefficient of your protein at 280 nm wavelength:**

Ext. coefficient: 29910 M-1 cm-1 Abs 0.1% (=1 g/l) 0.751, 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.

>gi|212659404:449825-453956 Toxoplasma gondii ME49 scf_1112359875116, whole genome shotgun sequence GAAAGGAAGACGGCAACATATTCGCGACGAATTCGGTGCATGCCTTCTAACTGGTTGTGTCTCTCCTCCC GTTCGTTAATACACTATGCTTGCCTCGGCCGAATGTTCTCAATCCCCCGCGGCCGCAGAACTGTTTCAAA ATAGAGAGACGAGACTAGAGCTGCGCGTCTCTTTTTGAGGCCGACCCGGTGTCATTTTTTTACAAGGGGA TTATGAAGTCGTGATTACAGCCTTTTTTTGACAAAGAGCTAAGGTCGCTGCTATGGTAATAAGAACGAAA ACCGTGTAAAGTGACCGGCGTTTCATCCTCCCTCTTGCGCTGACACAAAGTGACGCGGTGATCCAGCTTG ATTGAGTGACCTCTTCTTATTGTGCCTCTCTTTCAATTCATGCAAAAGCGTTCCCACGTCTGTACAGCCG TATCTGCATTCTGTTTTTGTGTCCCAGCCTGCTTCAAGCCAGTCAGTTGTGTAACGGTAAATGGAGGTGG CCGTTTCTTCCTCGGGACCTTCTTCCCTCCGCCCTGTCTCGGTCTACCACGGTGGAGCAAGCGGGTCTGA GGATAAGAAACTGTGGATCTGGCTACCGTACATTTCTGCATTATCGCGCAACGGCTTCGGCACCTCGAAT AATTCGCAGTTGGCATCCCGCGGCTCTCTGCGCCCCTCCGTGGCGTCAATGCAGAGTGCTTCTTCTGCCG TTGGAGGTTCACTACCAGGAGTATCTTCGTCGGTGAACACATCCGCGGATGAGACGTGGGAAACAGTACG CAATATGGTTGTTGGAAGGCCAGTGACTCTTCAGTCCGCGGAGCGAATCCGAGCAGGGTCAGCACCCGCT AGCAGTCTCATGCCGAAGTGTCCCACAGACATCTATCCTCCGGGTTTGGTACCCACTCTGACGAAAGGTA GCCTTTTTGTGGGGTCCCTGAAGCACGCACTCGACGAGAACCTTCTCCTAAAATTCAACGTGAAACTCGT TGTTACAGTTGCGTGGCCATACGGAAGTTGGCCGCTTCAACAGCGCGTGCTCTATTCTCGTCTTGGGATC AGCCACATCAACCACCCTCTCTTGGATTCTCCGTCCCAGGTGAGCTCAGCACACAGAAGTCGGTAACGTT GTGACCGTGGCGGACGGAGGCACACGCGACAGGGTGGAAACGGTGCGTTTGGTTAGACTTCACTTTTAGG AGGTAAAACCGCTTGACGTGGGCTCGCTAGCGGCTCGAGACAAATTTTGCACATGGACTCATGCAAACGG ACTTCCCTGTGAAATGAGACAGTGAACCGAGGATTGTAATGCGTTCTTTCAAGCCCAACTTGCTCGTGAA TAACGTCTTCAGTGTTAGTCTGGCAAGGATGTTGTACCGAATGAGATTGTGAACTCACGCTTGCTGGGAT TGGGTTCGCTGCGTTTTTGTTAGGAGCTCGACTTTTCCCGCCTCTCATTGGCACGTATTCACTCATACCT CGCCAGGTAAGTGGGCATCGACTGTATACCGCAATTCCTTGTTGGGTTTTCTTCTGTTCATTCATGGCCC AAATTTCAGGTTTCCGTCACGAAGTTGATAGGTTTTTCCCCTCATCAGAGCAGATGTGTACCTCCGGTTT CACCTGCAGGAACTACGATAAGCAGTTGCTTACTGGTGGACACAGGGAAATTGAATATGGACTCTGCCAG TGATGGTGTTAATCCTTTTCTATTAATCGTTCCCGCGGTCAATTTATAACATTCCCCGCATACGTTGTAG CACTTCTGCGTAAGACAGAACCACTACTGGCTCGTCTTCCGTGTCATGCCCGTGAAATCGTCGTTGTACT GTCGCATAGGAGTTGCTTTTTCTTTATCGAGTGACTAGACGCGATTCATCTGTGAGGCGTGGCATTTTAG CGAGTGATGTTGCACTTTTACACACATGCATGCAACTCACCCTTTTTAACCAAATGTGTGTATATGCGCG GGCACACGCATATGATGGCATTAGTACGTTGTGTGTATTAAAACGGGTTGTGAGTTATTGACGCGGCCCC CAACTGGGATGTGAAGGTGAAATTTTCGAGTGCGGCAAGCATTTTGAATGTGTTGCTCTTGTTTTTGCGC CGACTGCATCTCTTGCAGGGGAGTGACAGTGTTAGTTCACTGTGAAAAGGGCATCTCTCGGTCTGTCTCC CTCTGTGCGGTGAGCGAAGATCTGTTACAGCTTCCAAATCAACACCGTTCGAAGCCTTCTGCTCTTCTTA TGTGTAATAACTATCTACGTTAAGCTTTTTGCACTGCGAATAACTTGTATGCCTTGCCTGAAGAATACAC TTGTTAAGGCAGAAAAGCAGGTAATCTGGTCAGCAGTGGCACATGACGACAGCATATGCTGGACTCGGGT TCTTCCTTTCAATTGGAGAGGCACAGATGTGAGAACATACTGGCACATTGATGTGATTCGAGTACGTGTG GCTTTGTAGTGTCCTCAGAGTCCAGTTTGATCGAATAGGCATTCAAAACCTCGGAACACAGCTGGTGGAC TTCCAATGTGACAACGCGTTTGAATAATTTCTTTTCGACGTATGGCCTGTTGTATGGCAACAAATGGATT TCATGCGTGCGATTTTCATGGCAGGCATATCTCATCGTTTATCATGGGCACACAACCATGTCTGCGTTGG AGGCAATTCGTAAATATCGCCCCATTGCGCGGCCGAATCCCGGGTTTTTTGTGCAACTCCAACGACTAGA AAGCAATCATCACACGGTGACAGCGTCTTTTCAAAACAGCTCTAACTCGCCATCAAGGCGCTCCCCTTCG AGAGTCGGTACGGAATTATCTAGTGGACGCTGATTCGGGATCTGTGGTGGGAAAAGCACATATCCCTATG GCAGGAATAGAGGTCGTCTAACGGTTATCAGTGTATACACCTTTCGTACTTATGTGTTTGCTTTGTGTCT CTTCTTCATTTGCCCATAGACTGGCCCCACTTCCACTGCTTCCCCACCTTCTCCTTCCCCTGTCGTCCTG AAGGGTCCTGTGTTGGGTCCAAGGAGGAGCGTATAATGTTGATGCGTCAGGGAACAAAAGAGAAGGTGTT GCAGAGCGGTAAACGGGCAGAGATGGAGTTGTGAACGGTGACGTATGAGGGTCCAGCATCGATACACTCA ATGAGAGACTGGCACTGAGCACGGCATGGAACTGCCGCTTCAGACTATGCAATTATTTGCTGGTCACCAG TCGAAGCACTTGCTGGCCTCTGGTGAGTCTCCGGTGGATGCGATCGGCAGACGAATGCGAAGAAGACGGT GCTTCAAGCACGCGACATCCCGCGCAAGATGGTCCAGACACCGTGTGCTTCAGTGGCAATTCAGGTTTAA AAAGTACCGATCTTCAGATGTATAGCGCTGCACCGGAGTAATCCATGTATCGACGTCGTGCGACACTGCT AGCAGGAAAGGCAGTCGTATTCGCCAGCAAGTGCATCTCGTAACTCGTTGTGAACAGGTGGTTAAGTGTG ACGTCCATGAGAAATTGACACCATTTTTGCATTCTCGCCTTAGTTTTAGGTCGCAACTTCGATTCTATAT GCAAGAATTTTCCCATAGCAACGGTCACAGTGTGCCCGTCGAGTTCTGAGTGTCTTAACCCCGACGAGTT TAGTGTGGTACACGGGGCCGCCATGTCGGGGTATAGACCTACGGATCCTTAGCTGAAAAAATAGAAAAGA CGACGGTGTGCGTGTGTGTTGGCAGTTGAGCAGGTGGTTGTCGTTGCATATGCGCTGACTTATGCGCCGA ATGGTCAATGCGATGGGCCGGTGTCACTCGTTAATCCGTCTTTTGTGAAGCGGTCCAGAATTAAGCAGAA CTTTGTGTAAAGGATTTTGATAGTCGTCACCGCCGCCTATACACTGTAGGTATTTTTGTCCAGGCTTGTC GGTTCCGTAGGTTTCCCAATGAGGTGTTTCTAGCGTGAGAACTGCGTTCCAGTCGTGTTGCTTCACTTGG ATATGCCCCCCAACCACTGCCCGAGCAAAGTGCTGCTTGTCACGTGCGGTGGAAGCGTTTTTTCTCGAAA GGTAGATTGTCGTCGTTGTAAGGACCTGTTTTCGAGATTTTAGTGTTACGGGTTCAATTTTAGCGACTTT CG
 * CDS Gene Sequence (paste as text only):**


 * GC% Content for gene:** 7.14%


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


 * GC% Content for gene (codon optimized):**

Do Not Need this info for Spring (but still copy these lines to your Target page for now) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
 * 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):**

Resources:
See ** ProtocolTargetDiscoveryVDS.docx ** for more Etiologic Risk Group Categories (for pathogens): http://www.utexas.edu/research/rsc/ibc/agent_class.html#_Toc7238334

Enzyme Explorer @http://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer.html
 * SIGMA-ALDRICH RESOURCES **

Enzyme Classification Index (EC number) @http://www.sigmaaldrich.com/life-science/biochemicals/biochemical-products.html?TablePage=14573088

WolframAlpha @http://www.wolframalpha.com/ DrugBank @http://www.drugbank.ca/

@http://www.niaid.nih.gov/Pages/default.aspx @http://eupathdb.org/eupathdb/ @https://patricbrc.vbi.vt.edu/portal/portal/patric/Home @http://www.nmpdr.org/FIG/wiki/view.cgi/Main/EssentialGenes @http://tubic.tju.edu.cn/deg/ @http://csgid.org/csgid/cake/pages/community_request_gateway @http://tdrtargets.org/ @http://gsc.jcvi.org/status.shtml
 * Databases of genes/organisms: **

@http://wwwnc.cdc.gov/eid/pages/scientific-nomenclature.htm
 * Scientific Nomenclature page from Center for Disease Control (gene, protein names and abbreviations) **

NCBI GENE Page: @http://www.ncbi.nlm.nih.gov/gene BLAST Page: @http://blast.ncbi.nlm.nih.gov/
 * Gene Information: **

NCBI Protein Page: @http://www.ncbi.nlm.nih.gov/protein Protein Expression Website Protein Expression Paper:
 * Protein Information: **


 * Primer Overlap PCR Articles **


 * Is my target good for Virtual Screening programs? **