TargetF15+-+Protein+Phosphatase+2A,+Putative+(Trypanosoma+brucei)

 ***NCBI Gene # or RefSeq#:** 3656063 ***Protein ID (NP or XP #) or Wolbachia#:** XP_843718.1  **Etiologic Risk Group (see link below):** 2  ***/Disease Information (sort of like the Intro to your Mini Research Write up):** African Trypanosomiasis, colloquially known as the sleeping sickness, is a potentially fatal disease occurring in 36 sub-Sahara African nations. In 2012 6,314 new cases of Trypanosomiasis were reported to the WHO, although it is estimated that over 20,000 new cases evolved in 2012 and 65 million people are currently at risk for contracting the disease. Trypanosomiasis is caused by the infection of protozoan parasites: Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense. T. b. gambiense accounts for 98% of human infections, whereas T. b. rhodesiense is accounts for only 2% but is faster acting. Both Species are transmitted to humans via bites from the Tsetse fly of the genus Glossina. Sleeping sickness is categorized into two stages. During the haemo-lymphatic stage T. brucei multiplies in subcutaneous tissue. This first stage is characterized by minimal symptoms such as fever and itching or is asymptomatic. The meningo-encephalic stage is the final stage in which T. brucei invades the central nervous system resulting in sensory and sleep cycle disturbances. If left untreated, Trypanosomiasis is fatal. http://www.tdrtargets.org/published/browse/t/91 http://www.ncbi.nlm.nih.gov/protein/XP_843718.1 Is it a monomer or multimer as biological unit**? (make prediction at** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): Multimeric state 8  **Complex of proteins?:** No  **Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):** Druggable in Human, 2NPP chain F with Microsystine http://dx.doi.org/10.1016/j.cell.2006.11.033
 * *Target (protein/gene name): ** Protein Phosphatase 2A, Putative
 * *Organism (including strain): ** Trypanosoma brucei brucei (TREU927)
 * Link to TDR Targets page (if present): **
 * <span style="font-family: Arial,sans-serif; font-size: 10pt; line-height: 1.5;">Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **
 * <span style="font-family: Arial,sans-serif; font-size: 10pt; line-height: 1.5;">Essentiality of this protein: **significant loss of fitness in bloodstream form (3 days)

http://www.brenda-enzymes.org/enzyme.php?ecno=3.1.3.16
 * E.C. #:** 3.1.3.16
 * Link to BRENDA E.C. # page:**
 * BRENDA enzyme mechanism schematic :**

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2965627/
 * Enzyme Assay Information:**
 * Colorimetric assay of protein phosphatases using p-Nitrophenylphosphate
 * Colorimetric assay of protein phosphatases using a synthetic phospho-peptide substrate and malachite green reagent
 * Assay of protein phosphatases using 32P-labeled substrates
 * Link to paper with assay information, required reagents, and step-by-step protocols:**
 * Costs, quantities of substrate reagents, suppliers, and catalog #'s for the colorimetric assay of protein phosphatases using p-Nitrophenylphosphate**
 * 10 mM pNPP solution (solution can be made from tablets diluted with water). pNPP is sold in packages of 5 tablets for $34.10 per package. The supplier is Sigma-Aldrich and the catalog number is N1891-5SET.
 * 1x calorimetric buffer (see website above for full ingredients list).
 * 5 N NaOH. 50% sodium hydroxide solution is sold in bottles of 25 ml for $26.70 per bottle. The supplier is Sigma-Aldrich and the catalog number is 415413-25ML.

Homology Model: PDB# 2YNL of Human PP2A. Pairwise Alignment:
 * Structure:**

Query Coverage: 476 Max Identities: 71% % Positives: 84% Chain used for Homology: C __Okadaic Acid and Microcystin__: Have been used to discover the function of PP2A in cells, but are too toxic to be used as drugs __Fostriecim__: Causes apoptosis of cells by inducing early mitosis; is highly susceptible to oxidation and Phase I trials were cancelled; is currently being tested for ways to prevent its oxidation __Canthardin/Norcanthardin__: Canthardin is a strong inhibitor with an IC50 of 0.194 and it's able to transverse membranes; its analog Norcanthardin is currently being tested as a drug candidate because Canthardin has high toxicity levels Expressed in //E. coli// cells: http://www.sciencedirect.com/science/article/pii/S0378111901003675 Similar Protein PP5 was purified with Ni-NTA chromatography using 6His tagged proteins: http://www.sciencedirect.com/science/article/pii/S0378111901003675
 * Current Inhibitors:**
 * Expression Information:**
 * Purification Method:**
 * PyMol Image of Protein:**

<span style="font-family: Arial,sans-serif; font-size: 9pt;">MPATTSIPPVTAPGTGGEASSFNVDELIQYVLQGKPLSEPQVARLCQKAREVLEKEENVHTVRAPVTVCGDIHGQFHDLLELLKIGGLPPDTNYLFMGDYVDRGYYSVETVTLLLLFKIRY <span style="font-family: Arial,sans-serif; font-size: 9pt;">PERVQILRGNHESRQITQVYGFYDECVRKYGSANVWKLLTDLFDYLPLAAVVENEIFCLHGGLSPTLDSLAHIRSLERVQEVPHEGPMCDLLWSDPEDKDGWGISPRGAGFTFGADIT  <span style="font-family: Arial,sans-serif; font-size: 9pt;">EQFCHNNGLKTIARAHQLVAEGYSWAHSDKLVTIFSAPNYCYRCGNLAGLLELDEHMNKCFFQFDPAPRRGEAQVSKKTPDYFL
 * Amino Acid Sequence:**


 * Length of protein in Amino Acids:** 323 aa


 * Molecular Weight of protein in (kD):** 36,243.1

Ext. coefficient 43360 Abs 0.1% (=1 g/l) 1.197, assuming that all pairs of Cys residues form cystines Ext. coefficient 42860 Abs 0.1% (=1 g/l) 1.183, assuming that all Cys residues are reduced
 * Molar Extinction coefficient at 280 nm wavelength:**

TMpred graph:
 * [[image:sneeeeep.PNG width="408" height="306"]]

Most likely no transmembrane proteins; no possible models found.

<span style="font-family: Arial,sans-serif; font-size: 10pt;">ATGCCGGCGACCACCTCTATCCCGCCTGTGACTGCGCCTGGTACCGGTGGTGAGGCGTCT TCCTTCAATGTTGACGAGCTCATCCAGTACGTTCTGCAGGGTAAACCG CTGTCTGAACCGCAGGTTGCCCGTCTGTGCCAGAAAGCGCGTGAAGTTCTGGAAAAAGAAGAAAACGTGCACACCGTTCGTGCGCCAGTTACCGTTTGCGGTGACAT CCATGGTCAGTTCCACGATCTGCTGGAACTCCTGAAAATTGGTGGTCTCCCACCGGACACCAACTATCTCTTCATGGGTGACTACGTTGATCGCGGCTACTACTCCGTTG AAACCGTTACGCTCCTCCTGCTGTTCAAAATCCGCTACCCGGAGCGTGTTCAGATCCTGCGTGGTAATCATGAATCTCGTCAGATCACCCAGGTTTACGGCTTCTACGAC GAATGCGTTCGTAAATACGGTTCTGCGAATGTTTGGAAACTGCTGACGGATCTGTTCGACTACCTGCCGCTGGCGGCTGTTGTTGAAAACGAAATCTTCTGCCTGCACGG TGGCCTGTCTCCGACCCTGGACTCTCTGGCTCACATCCGTTCTCTGGAACGTGTCCAAGAGGTCCCGCATGAAGGCCCGATGTGCGACCTCCTCTGGTCCGATCCGGA AGACAAAGACGGTTGGGGTATCTCTCCGCGTGGTGCGGGTTTCACCTTCGGTGCGGACATTACCGAACAGTTCTGCCACAACAATGGTCTGAAAACCATCGCGCGTGC TCACCAGCTCGTAGCCGAGGGTTACTCTTGGGCGCACTCTGATAAACTGGTTACTATCTTCTCTGCGCCGAACTACTGCTATCGTTGTGGTAACCTGGCGGGTCTGCTCG AACTCGACGAACACATGAACAAATGCTTCTTTCAGTTCGACCCGGCACCTCGTCGCGGTGAAGCACAAGTCTCCAAGAAGACCCCGGACTACTTCCTGTAA
 * CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):**
 * GC% Content for gene (codon optimized):** 54.7%