Target+-+Inorganic+pyrophosphatase+(T.+brucei)....................................................Shannon+R.

** *Target (protein/gene name): Inorganic pyrophosphatase **
This protein was found to be essential in inhibiting T. brucei during an experiment done with mice (see article).
 * *NCBI Gene # or RefSeq#: 3656220 (Gene #) **
 * *Protein ID (NP or XP #) or Wolbachia#: 15644 **
 * *Organism (including strain): **** Trypanosoma brucei **
 * Etiologic Risk Group (see link below): Parasitic Agents **
 * *Background/Disease Information (sort of like the Intro to your Mini Research Write up): Trypanosoma brucei is a protozoan that causes African trypanosomiasis (sleeping sickness). This disease can be found in humans or animals. It is transmitted by the tsetse fly. African trypanosomiasis can be treated with medication. **
 * Essentiality of this protein: ** []
 * Complex of proteins?: **
 * Druggable Target: No **


 * *EC#: 3.6.1.1 **
 * Link to BRENDA EC# page: ** []
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic

[]
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * -- link to Sigma (or other company) page for assay or assay reagents (substrates) **

[]
 * -- link (or citation) to paper that contains assay information **

$237.00 from Sigma Aldrich -- PDB # or closest PDB entry if using homology model: 3EMJ -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB
 * -- List cost and quantity of substrate reagents and supplier **
 * Structure Available (PDB or Homology model) **

Query 74 RRTRAKLELVKEEPHN--PIAQDTLKKEGNAL---RFFKYG-DVPFNYGFAPQTWEDPSV 127 + ++ ++ E P N PI + KE AL RF + P NYGF P T Sbjct 10 KANNNEINVIIEIPMNSGPIKYE-FDKESGALFVDRFMQTTMSYPCNYGFIPDTL- 63

Query 128 MDQLTTCGGDGDPIDIVELSSNPFAVGSVRAVRVLGLLGLIDEGETDWKVITEAIGP-DA 186 DGDP+D++ ++ +P GSV R +G+L + DE D K+I D Sbjct 64 ---SNDGDPVDVLVVAHHPVVPGSVIKCRAIGVLMMEDESGLDEKIIAVPTSKLDI 116

Query 187 TGTY-GSLNNVPQELKATIVKWFREYK 212 T + L+++ + LK IV +F YK Sbjct 117 TFDHIKELDDLCEMLKKRIVHFFEHYK 143

Query Coverage: 56% Max % Identities: 56% % Positives: 55% Chain used for homology: 3EMJ

mrptsimfkgmtgagimlpawalqevgaagtrawrmyftsseagsvarrsawhdlplhps pdasvitfvceiprrtraklelvkeephnpiaqdtlkkegnalrffkygdvpfnygfapq twedpsvmdqlttcggdgdpidivelssnpfavgsvravrvlgllglidegetdwkvite aigpdatgtygslnnvpqelkativkwfreyktadgkkpnefvfggelrnaddalrvieg gsrqytgliagtvrnpgywlh >gi|72386904|ref|XM_838784.1| Trypanosoma brucei inorganic pyrophosphatase, putative (Tb927.3.2840) partial mRNA ATGCGCCCAACATCAATTATGTTTAAAGGGATGACCGGAGCAGGCATTATGCTCCCCGCATGGGCATTGC AGGAAGTGGGAGCAGCGGGAACGCGGGCGTGGCGCATGTACTTCACAAGCAGCGAGGCCGGATCCGTTGC GAGGCGCTCGGCATGGCATGATCTTCCGCTACATCCATCACCTGATGCATCTGTCATTACATTTGTATGT GAGATTCCCAGAAGGACGCGTGCCAAATTGGAACTCGTGAAGGAAGAACCGCACAACCCGATAGCTCAAG ACACATTAAAGAAAGAGGGCAATGCTTTGCGGTTCTTCAAGTATGGTGATGTCCCCTTCAACTATGGTTT TGCGCCACAAACATGGGAGGATCCTTCAGTAATGGACCAACTGACGACGTGTGGGGGTGATGGTGATCCC ATTGATATCGTTGAATTATCGTCAAATCCCTTTGCAGTTGGTTCCGTAAGGGCAGTTCGAGTCCTCGGGT TGTTGGGACTTATTGACGAGGGGGAAACCGACTGGAAAGTCATTACGGAGGCCATTGGACCCGATGCGAC CGGTACGTACGGGTCTTTAAATAATGTGCCACAAGAGCTGAAGGCTACCATTGTTAAATGGTTCCGCGAG TACAAAACTGCTGATGGGAAGAAGCCAAATGAATTTGTTTTTGGAGGGGAGTTGCGTAATGCAGACGACG CACTTCGTGTGATTGAAGGAGGTTCCCGCCAGTACACCGGACTTATTGCAGGCACGGTTCGTAATCCCGG TTACTGGCTTCACTGA
 * Current Inhibitors: about 81 current inhibitors **
 * Expression Information (has it been expressed in bacterial cells): Yes it can be expressed in E.coli **
 * Purification Method: Recombinant protein using His-TAG **
 * Image of protein (PyMol with features delineated and shown separately): (Image of inorganic phosphatase for Rickettsia prowazekii due to unavailability of protein in Trypanosoma brucei) **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **
 * *length of your protein in Amino Acids: 261 **
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: 28675.5 **
 * Molar Extinction coefficient of your protein at 280 nm wavelength: 49055 **
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. [[image:vdsstream/TMPRED.17150.8689.gif width="448" height="336" link="@http://www.ch.embnet.org/software/TMPRED_form.html"]]
 * *CDS Gene Sequence (paste as text only): **
 * *GC% Content for gene: **
 * *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

@http://www.niaid.nih.gov/Pages/default.aspx @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: **

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: **