TargetSp15+-+2-acetamido-2-deoxy-alpha-D-glucopyranosyl-(1->6)-phosphatidylinositol+de-N-acetylase+(Trypanosoma+brucei)


 * Target (protein/gene name)**: 2-acetamido-2-deoxy-alpha-D-glucopyranosyl-(1->6)-phosphatidylinositol de-N-acetylase


 * NCBI Gene #** : 719226


 * Protein ID**: fig|870187.4.peg.785


 * Organism**: Trypanosoma brucei


 * Etiologic Risk Group**: Risk group 2- Parasitic agents


 * Link to TDR targets page**: N/A


 * Link to Gene database (Patric):** https://www.patricbrc.org/portal/portal/patric/Feature?cType=feature&cId=PATRIC.870187.4.AJUL01000009.CDS.733719.734555.rev

According to the Brenda site, "N-ureido analogue, synthesised by us, had inhibitory activity against the aforementioned de-N-acetylase, presumably via the N-ureido motif."
 * Essentiality Info:**


 * Is it a Monomer or Multimer?**: monomer


 * Complex of Proteins**: N/A

http://www.brenda-enzymes.org/literature.php?e=3.5.1.89&r=649652
 * Druggable target:** The following info was found on Brenda: "Substrate specificity of the N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase of glycosylphosphatidylinositol membrane anchor biosynthesis in African trypanosomes and human cells."


 * EC** 3.5.1.89

Sleeping sickness, African trypanosomiasis, is a deadly blood disease caused by two variates of //Trypanosoma brucei// and transmitted by tsetse fly. In 1–3 weeks after the bite a chancre (a red sore skin lesion) can develop on the bite area. Several weeks or months later //Trypanosoma// parasites in the blood, spinal and lymphatic fluid (hemolymphatic stage) can cause fever, anemia, etc. As the disease reaches its final stage, the parasites get through the blood-brain barrier entering the brain. This meningoencephalitic stage (inflammation of the central nervous system) could cause blackouts, coma, or death.
 * Disease information**

Pentamidine isethionate and suramin are usually used for treating the hemolymphatic stage of West and East African Trypanosomiasis, respectively. Melarsoprol is used for late disease with central nervous system involvement. Tsetse flies are common only in rural Africa, not in urban cities, so this disease is considered an "Old World" disease.

http://www.brenda-enzymes.org/enzyme.php?ecno=3.5.1.89
 * link to Brenda EC page**:

Figure 1: Reaction schematic on Brenda site

http://www.ncbi.nlm.nih.gov/pubmed/20085486
 * Enzyme Assay Info**: non-radioactive high-throughput assay

6-(N-acetyl-alpha-D-glucosaminyl)-1-phophatidyl-1D-myo-inositol: no info found water: found in lab currently
 * List cost and quantity of substrate reagents, supplier, and catalog #**


 * Structure:** N/A

http://www.ncbi.nlm.nih.gov/pubmed/18479678
 * Current inihibitors:** "Synthesis of 1-D-6-O-[2-(N-hydroxyaminocarbonyl)amino-2-deoxy-alpha-D-glucopyranosyl]-myo-inositol 1-(n-octadecyl phosphate): a potential metalloenzyme inhibitor of glycosylphosphatidylinositol biosynthesis"


 * Expression Information**: No information was found on expression in other bacteria.


 * Purification method**: E-52 cellulose batch preparation followed by 2' AMP-agarose affinity chromatography


 * Image in Pymol**:

0 MCPINASFYT LIVVKLLRES LIHMHGALAF GFVVVFLSFL VLWQRASCVS KIHLVGDVLF 60 VFAHPDDEAM FFSPLLDYVR RHGLNAHFLC LSNGNYSGLG TVREKELVAS AEYFGVNRRS 120 VRVVDHPDLQ DGPDNLWNTE IVQREVLSYL HSVKDIRTVI TFDHRGVSSH ANHVAVYEGV 180 LLAKKNLPPG ILFLSLHTRD LLEKYVGILS TVGYTVGIHR CGGRRNHVIL IPPTSLFTSF 240 SAMRKHKTQL VWFRYLFVWF SSYSYVNEVK ELGVA
 * Amino Acid Sequence**:


 * Length of Protein:** 275


 * Molecular Weight:** 31205 kD


 * Molar Extinction coefficient of your protein at 280 nm wavelength:** 38640


 * TMpred graph Image ([]):**

Figure 2: TMpred graph of 2-acetamido-2-deoxy-alpha-D-glucopyranosyl-(1->6)-phosphatidylinositol de-N-acetylase of T. brucei run using amino acid sequence.

http://www.bioinformatics.org/sms2/rev_trans.html
 * CDS Gene Sequence (paste as text only): **

atgtgcccgattaacgcgagcttttataccctgattgtggtgaaactgctgcgcgaaagc ctgattcatatgcatggcgcgctggcgtttggctttgtggtggtgtttctgagctttctg gtgctgtggcagcgcgcgagctgcgtgagcaaaattcatctggtgggcgatgtgctgttt gtgtttgcgcatccggatgatgaagcgatgttttttagcccgctgctggattatgtgcgc cgccatggcctgaacgcgcattttctgtgcctgagcaacggcaactatagcggcctgggc accgtgcgcgaaaaagaactggtggcgagcgcggaatattttggcgtgaaccgccgcagc gtgcgcgtggtggatcatccggatctgcaggatggcccggataacctgtggaacaccgaa attgtgcagcgcgaagtgctgagctatctgcatagcgtgaaagatattcgcaccgtgatt acctttgatcatcgcggcgtgagcagccatgcgaaccatgtggcggtgtatgaaggcgtg ctgctggcgaaaaaaaacctgccgccgggcattctgtttctgagcctgcatacccgcgat ctgctggaaaaatatgtgggcattctgagcaccgtgggctataccgtgggcattcatcgc tgcggcggccgccgcaaccatgtgattctgattccgccgaccagcctgtttaccagcttt agcgcgatgcgcaaacataaaacccagctggtgtggtttcgctatctgtttgtgtggttt agcagctatagctatgtgaacgaagtgaaagaactgggcgtggcg


 * GC% Content for gene**: 55.03%


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


 * GC% Content for gene (codon optimized): N/a**

(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- 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):**
 * Primer design results for 'tail' primers (this is just 2 sequences):**