TargetSp15+-+Phosphotyrosine+protein+phosphatase+(Mycobacterium+tuberculosis)

M. tuberculosis causes tuberculosis, a potentially fatal infectious pulmonary disease. Tuberculosis can be spread through the air through small droplets containing the bacteria from the coughs of sneezes of infected persons. Tuberculosis has become more common in developed countries due to the spread of HIV, which makes patients more susceptible to tuberculosis. Tuberculosis occurs in both a latent and active form and results in symptoms including coughing up blood, chest pain, weight loss, fatigue, fever, chills, and loss of appetite. Especially common in low and middle income countries, tuberculosis has the second highest worldwide mortality count for a single infectious agent. Treatment for tuberculosis currently requires multiple drugs for at least six months and without proper treatment, up to 66% of patients will die. Homo sapiens –“MptpB is a promising target for development of anti-tuberculosis drugs” - http://www.brenda-enzymes.org/literature.php?e=3.1.3.48&r=710432 Macaca fascicularis - “PTP-1B antagonizes insulin signaling and is a potential therapeutic target for insulin resistance associated with obesity and type 2 diabetes” - http://www.brenda-enzymes.org/literature.php?e=3.1.3.48&r=692092
 * *Target (protein/gene name): **Phosphotyrosine protein phosphatase (ptpB)
 * *NCBI Gene # or RefSeq#: **886842
 * *Protein ID (NP or XP #) or Wolbachia#: **NP_214667
 * *Organism (including strain): **Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
 * Etiologic Risk Group (see link below): **Risk Group 3
 * * Disease Information (sort of like the Intro to your Mini __Research Write__ up): **
 * Link to TDR Targets page (if present): ** Not present
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): **@http://www.ncbi.nlm.nih.gov/gene/886842
 * Essentiality of this protein: **“essential virulence factor that is secreted by the bacterium into the cytoplasm of macrophages, where it mediates mycobacterial survival in the host” (@https://www.ncbi.nlm.nih.gov/pubmed/20167798)
 * Is it a monomer or multimer as biological unit? ** : Monomer
 * Complex of proteins?: **One of three phosphatases (PtpA, PtpB, and SapM)
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **

https://www.neb.com/products/p0757-p-nitrophenyl-phosphate-pnpp http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3097125/ -NaOH (Sigma Aldrich, #221465, $36.00 for 25g) http://www.sigmaaldrich.com/catalog/product/sial/221465?lang=en®ion=US -pNPP (Sigma Aldrich, #N7653, $89.60 per pack) http://www.sigmaaldrich.com/catalog/product/sigma/n7653?lang=en®ion=US -Alkaline Phosphatase Stabilizing Buffer (Sigma Aldrich, #A4955, $92.70 for 100ml) http://www.sigmaaldrich.com/catalog/product/sigma/a4955?lang=en®ion=US
 * *EC#: ** 3.1.3.48
 * Link to BRENDA EC# page: ** @http://www.brenda-enzymes.info/enzyme.php?ecno=3.1.3.48
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **continuous spectrophotometric assay
 * -- link to Sigma (or other company ) page for assay (see Sigma links below) **
 * -- links to assay reagents (substrates) pages; **** List cost and quantity of substrate reagents, supplier, and catalog # **

-- PDB #: 1YWF
 * Structure (PDB or Homology model) **

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2775457/ http://www.biomedcentral.com/1471-2180/13/292 MAVRELPGAWNFRDVADTATALRPGRLFRSSELSRLDDAGRATLRRLGITDVADLRSSREVARRGPGRVP DGIDVHLLPFPDLADDDADDSAPHETAFKRLLTNDGSNGESGESSQSINDAATRYMTDEYRQFPTRNGAQ RALHRVVTLLAAGRPVLTHCFAGKDRTGFVVALVLEAVGLDRDVIVADYLRSNDSVPQLRARISEMIQQR FDTELAPEVVTFTKARLSDGVLGVRAEYLAAARQTIDETYGSLGGYLRDAGISQATVNRMRGVLLG 30.1538 kDa atggcggtgcgcgaactgccgggcgcgtggaactttcgcgatgtggcggataccgcgacc gcgctgcgcccgggccgcctgtttcgcagcagcgaactgagccgcctggatgatgcgggc cgcgcgaccctgcgccgcctgggcattaccgatgtggcggatctgcgcagcagccgcgaa gtggcgcgccgcggcccgggccgcgtgccggatggcattgatgtgcatctgctgccgttt ccggatctggcggatgatgatgcggatgatagcgcgccgcatgaaaccgcgtttaaacgc ctgctgaccaacgatggcagcaacggcgaaagcggcgaaagcagccagagcattaacgat gcggcgacccgctatatgaccgatgaatatcgccagtttccgacccgcaacggcgcgcag cgcgcgctgcatcgcgtggtgaccctgctggcggcgggccgcccggtgctgacccattgc tttgcgggcaaagatcgcaccggctttgtggtggcgctggtgctggaagcggtgggcctg gatcgcgatgtgattgtggcggattatctgcgcagcaacgatagcgtgccgcagctgcgc gcgcgcattagcgaaatgattcagcagcgctttgataccgaactggcgccggaagtggtg acctttaccaaagcgcgcctgagcgatggcgtgctgggcgtgcgcgcggaatatctggcg gcggcgcgccagaccattgatgaaacctatggcagcctgggcggctatctgcgcgatgcg ggcattagccaggcgaccgtgaaccgcatgcgcggcgtgctgctgggc
 * Current Inhibitors: **I-A09 (https://www.ncbi.nlm.nih.gov/pubmed/20167798)
 * Expression Information (has it been expressed in bacterial cells): ** Expressed in e.Coli
 * Purification Method : ** immobilized metal affinity chromatography, and size exclusion chromatography (can also use a cobalt charged resin column)
 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **
 * *length of your protein in Amino Acids: **276aa
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: **
 * Molar Extinction coefficient of your protein at 280 nm wavelength: 14440 **
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
 * *CDS Gene Sequence (paste as text only): **
 * *GC% Content for gene: **65.821%
 * *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): **