TargetSp17-+PP2C-+family+Ser+Thr+Phosphatase+(Mycobacterium+tuberculosis)


 * Target (protein/gene name): PP2C- family Ser ****Thr Phosphatase (Mycobacterium tuberculosis)**


 * *NCBI Gene # or RefSeq#: 887070 **


 * *Protein ID (NP or XP #) or Wolbachia#: AF177866_1 **

//Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv) //
 * *Organism (including strain): **

N/A Tuberculosis is a contagious bacterial infection is spread through the air. Tuberculosis usually attacks the lungs and is caused by a bacteria called Mycobacterium tuberculosis. The symptoms of Tuberculosis include cough, coughing up blood, chest pain and fatigue. If the Tuberculosis infection worsens or goes untreated, it can infect the central nervous system, kidneys, lungs, heart and brain.
 * Etiologic Risk Group (see link below): **
 * /Disease Information (sort of like the Intro to your Mini Research Write up):**

N/A
 * Link to TDR Targets page (if present): **

[] The only predicted protein phosphatase in M.tuberculosis, it dephosphorylates at least 5 protein kinases (PknA, PknB, PknD, PknE and PknF) and the penicillin-binding protein PBPA - Dr. B 060617 Is it a monomer or multimer as biological unit ** ? (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): Ortholog of A. thaliana: monomer
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **
 * Essentiality of this protein: **


 * Complex of proteins?: ** No

http://www.ebi.ac.uk/enzymeportal/search/P9WHW5/enzyme
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **

http://www.rcsb.org/pdb/explore.do?structureId=1TXO
 * Link to PDB:**


 * *EC#: ** 3.1.3.16

http://www.brenda-enzymes.org/enzyme.php?ecno=3.1.3.16&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Mycobacterium+tuberculosis&show_tm=0
 * Link to BRENDA EC# page: **

The purified wild-type or mutant proteins were exchanged into reaction buffer containing 50 mM NaCl, 20 mM Tris, pH 7.5, and 0.5 mM TCEP. The enzymes were incubated with various amounts of pNPP, and reaction progress data were collected at 405 nM and 25°C using a SPECTRAmax 190 spectrophotometer (Molecular Devices). Kinetic constants were calculated using SigmaPlot (Systat Software Inc.). In order to determine the effect of metal concentration on enzyme activity, various amounts of MnCl 2 were mixed with 50 mM pNPP. The reaction was initiated by adding enzyme at a final concentration of 1 μM, and the assay was monitored continuously for several minutes. To determine the k cat and K m values for pNPP, eight substrate dilutions were made. Reactions contained 100 mM MnCl 2 and a final enzyme concentration of 1 μM.
 * Enzyme Assay information (spectrophotometric, coupled assay ? **

http://www.sciencedirect.com/science/article/pii/S096921260400334X

http://www.sciencedirect.com/science/article/pii/S0006291X03020011?via%3Dihub

@http://www.uvm.edu/~bio1and2/lab/Lab%20manuals%20Fall%202011/Enzyme%20Activity%20Phosphatase.pdf
 * -- 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 # **

-- PDB # or closest PDB entry if using homology model: -- 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 (PDB or Homology model) **



recombinant wild-type and selenomethionine-labeled enzymes from Escherichia coli strain BL21(DE3) by gel filtration and ion exchange chromatography code >AAF04553.1 putative protein phosphatase type 2C, partial [Caenorhabditis brenneri] SKVSQYSGINLHKKVVARKEFSEGNLKGAIERGFLDLDQQMRIDEETKDDVSGTTAVVVLIKEGDVYCGN AGDSRAVSSVVGEARPLSFDHKPSHENEARRIIAAGGWVEFNRVNGNLALSRALGDFAFKNCDTKPAEEQ IVTAYPDVITDKLTPDHEFIVL code 162 aa
 * Current Inhibitors: Phosphoserine, phosphothreonine, para-nitrophenol phosphate **
 * Expression Information (has it been expressed in bacterial cells): **
 * Purification Method:**
 * 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 **
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: 17762.90 kDa**
 * Molar Extinction coefficient of your protein at 280 nm wavelength: 10095**
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.



code >NC_000962.3:c23181-21637 Mycobacterium tuberculosis H37Rv, complete genome GTGGCGCGCGTGACCCTGGTCCTGCGATACGCGGCGCGCAGCGATCGCGGCTTGGTACGCGCCAACAACG AAGACTCGGTCTACGCTGGGGCACGGCTATTGGCCCTGGCCGACGGCATGGGTGGGCATGCGGCCGGCGA GGTGGCGTCCCAGTTGGTGATTGCCGCATTGGCCCATCTCGATGACGACGAGCCCGGTGGCGATCTGCTG GCCAAGCTGGATGCCGCGGTGCGCGCCGGCAACTCGGCTATCGCAGCGCAAGTCGAGATGGAGCCCGATC TCGAAGGCATGGGTACCACGCTCACCGCAATCCTGTTCGCGGGCAACCGGCTCGGCCTGGTGCATATCGG TGACTCGCGCGGTTACCTGCTGCGCGACGGTGAGCTGACGCAGATCACCAAGGACGACACGTTTGTCCAA ACGCTGGTCGACGAAGGCCGGATCACCCCGGAGGAGGCGCACAGCCACCCGCAACGCTCGTTGATCATGC GGGCGTTGACCGGCCATGAGGTCGAACCGACGCTGACCATGCGAGAAGCCCGCGCCGGTGATCGTTACCT GCTGTGCTCGGACGGGTTGTCCGATCCGGTTAGCGATGAAACTATCCTCGAGGCCCTGCAGATCCCCGAG GTTGCCGAGAGCGCTCACCGCCTCATTGAACTGGCGCTGCGCGGCGGCGGCCCCGACAACGTCACTGTCG TCGTCGCCGACGTCGTCGACTACGACTACGGCCAGACCCAACCGATTCTGGCCGGGGCGGTCTCAGGCGA CGACGACCAACTGACCCTGCCCAACACCGCCGCCGGCCGGGCCTCTGCCATCAGCCAGCGCAAGGAGATC GTTAAACGCGTTCCGCCACAGGCCGATACATTCAGTCGGCCACGGTGGTCGGGCCGACGGCTAGCATTCG TTGTCGCACTGGTGACCGTGCTGATGACTGCGGGCCTGCTCATTGGTCGCGCGATCATCCGCAGCAACTA CTACGTAGCGGACTACGCCGGCAGCGTGTCCATCATGCGGGGGATTCAAGGGTCGCTACTGGGCATGTCC CTGCACCAGCCTTACCTGATGGGCTGCCTCAGCCCGCGTAACGAGCTGTCGCAGATCAGCTACGGACAGT CTGGGGGCCCTCTCGACTGCCATCTGATGAAACTGGAGGATCTGCGACCGCCGGAGCGCGCACAGGTTCG GGCCGGTCTCCCGGCCGGCACTCTCGATGACGCCATCGGGCAGTTGCGCGAACTGGCGGCCAACTCCCTG CTGCCGCCTTGCCCGGCGCCGCGTGCCACGTCCCCGCCCGGGCGCCCGGCCCCACCCACCACCAGCGAGA CAACCGAACCAAACGTCACCTCCTCGCCAGCCTCTCCATCACCCACCACCTCCGCGCCGGCCCCCACCGG AACTACTCCTGCCATCCCCACGAGTGCCTCCCCGGCAGCGCCCGCGTCGCCGCCGACGCCTTGGCCCGTC ACCAGCTCGCCGACGATGGCCGCACTTCCGCCACCCCCGCCTCAGCCGGGCATCGACTGCCGGGCGGCGG CATGA code
 * CDS Gene Sequence (paste as text****only):**
 * *GC% Content for gene: **
 * 67.572816**
 * 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)

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

Reverse Primer:
TATCCACCTTTACTG TTA GTGTTCGAGGTCCGCAA **