*Target (protein/gene name): 6-phosphoglucanate dehydrogenase, decarboxylating, putative

*NCBI Gene # or RefSeq#: XP_808031.1

Gene ID: 3550073

*Protein ID (NP or XP #) or Wolbachia#: XP_817969.1

*Organism (including strain): Trypanosoma cruzi/ strain CL Brener

Etiologic Risk Group (see link below):

*/Disease Information (sort of like the Intro to your Mini Research Write up):
Over 6 million people living mostly in Latin American countries located in Central and South America are infected with Trypansosoma cruzi and have developed the illness known as Chagas disease, or American trypansosomiasis. It is recognized by the World Health Organization as one of the world’s 13 most neglected tropical diseases. Throughout the years, the disease has spread to other parts of the world, such as Europe, North America, and Asia due to high immigration and mobility between countries. The triatomine bugs, known as “kissing bugs”, are the carriers that transmit the parasite into the human population. Specifically, it is transmitted when the person rubs the urine or feces into the entrance of their skin, such as the bite area, mouth, and eyes. Infection can present itself through symptoms such as fever, nausea, and vomiting. These symptoms may go untreated resulting in the chronic stages of the infection. In the chronically ill, the disease can cause complications, such as life threatening heart disease and digestive disorders which can lead to heart failure, cardiomyopathy, megaoesophagus, and inflammation of the colon. The most common treatment for the early stages of the infection is the use of drugs called benznidazole or nifurtimox. They are ineffective during the late stages of the infection and have debilitating side effects. There have been growing resistance against the drugs. At the moment, there are no vaccines to prevent the infection, but a new study has shown a promising vaccine against the infection as seen through mice trials. Therefore, a new therapeutic approach is to target an essential protein, 6-phosphogluconate dehydrogenase (6PGDH), with

Link to TDR Targets page (if present): http://tdrtargets.org/targets/view?gene_id=49268

Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)
__https://www.ncbi.nlm.nih.gov/gene/?term=1.1.1.44+cruzi__
__http://www.uniprot.org/uniprot/Q6WAT4__


Essentiality of this protein:

Essential to the organism’s survival; main enzyme that participates in the pentose phosphate pathway. Responsible for the synthesis of nucleotides, amino acid production, fatty acid synthesis, and provides NADPH for reactions in the organism.
Is it a monomer or multimer as biological unit? (make prediction at http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html):

Complex of proteins?: 2 mer; two amino acid chain

Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
0.6 from TDR Targets

*EC#: 1.1.1.44
Link to BRENDA EC# page: __http://www.brenda-enzymes.info/enzyme.php?ecno=1.1.1.44__
-- Show screenshot of BRENDA enzyme mechanism schematic
external image F2RnfDRKNTZQtTD7HPAoeUqdAo8yIJtDvgqBUsb_JswpgVfefFe3liXS6GuMnXDN1LeD-TxIMdlXUhKvlbgre7qTK_p8hnFTxEOChyIXgsvuVCbkt3toulknPHKuQq2OjQ=s1600
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
Spectrophotometric → UV Quantification
-- 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.
__http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/6phosphoglucdehydro74.pdf__ (pH 7.4)
__http://www.sigmaaldrich.com/catalog/product/sigma/mak038?lang=en&region=US__
--- List cost and quantity of substrate reagents, supplier, and catalog #

Structure (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 1PGJ
-- For Homology Model option:
external image DiBVsOrkwOZbOUvbJwj_iM0cruP93Ty7HNo1Ln-Wv7IVJODdAelh0gRKTlBs1e_iYMGHFuk_uBsw4TnktyCCVjmGo6LrrHMGVvlHqqS_1U0OV-A8JNkrfh7YSr36s812=s1600


---- Query Coverage: 75%
---- Max % Identities: 100%
---- % Positives: 89%
---- Chain used for homology: A

Current Inhibitors: There are several inhibitors that have been tested against different compounds for T. cruzi and T. Brucei, but they seemed to work better for the T.Brucei.
Expression Information (has it been expressed in bacterial cells):

__http://www.scielo.br/pdf/aabc/v79n4/a07v79n4__
__http://www.ncbi.nlm.nih.gov/pubmed/14698432__

Purification Method: Nickel affinity chromatography
Image of protein (PyMol with features delineated and shown separately):
external image I-Dquz_vXmROpnYHYR7qPaEcTGaocC5sPRerDUDwf9LgkSBSe2su44zJYzaKi4498gJeznhht38OAQW_LwvdDB2nPejJnM8Eflg6obXhYaVLOiQi8iuX4KtYvpuA8lXFSw=s1600
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MDVGIVGLGVMGANLALNIAEKGFHVAVFNRTYSKAESFIKEHASAPFAANLKAYETMKS
FIMAIKKPRKIFILVQAGAATDATIEQLKGVLEKDDIIVDTGNAHFKDQTRRAEQLEAAG
LRFLGMGISGGEEGARKGPAFFPGGTLSVWKEIQPVVEAAAAKADDGRPCVTMNGRGGAG
SCVKMYHNAGEYAVLQVWGEAFDVLRAMGLSNDEVAAVFEDWKAKGFLTSYMLDISIVAA
RAKVADGSHLSEHVKDCIGSKGTGLWSAQEALEVGVPAPSLNMAVLSRQMTMYKSERELN
AKTLPFVLKVPGYAIKDKSPNAPEIRQLYHAVSIAIIACYAQMFQCLRALDKVYDFGLDL
PATIATFRAGCILKGYLLQPMTEAFAKNVKLSNLLCAFEKEIREGLQSYRDILGFITSKT
ALTLPVLSASLVYVNGMFTPTLRYGQLVSLQRDVFGRHGYERLDKEGRESHQWPELQ
*length of your protein in Amino Acids: 477 aa
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website:
5,2134.4 Da
Molar Extinction coefficient of your protein at 280 nm wavelength:

Ext. coefficient50225
Abs 0.1% (=1 g/l) 0.963, assuming all pairs of Cys residues form cystines

Ext. coefficient49850
Abs 0.1% (=1 g/l) 0.956, assuming all Cys residues are reduced

TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
external image kfdW4hfaC14M1TQQKBaDm9-hzrMRabQ_eMDj0LCkrK3e3UuGTMAWIHVlDfFD0FX1johXNytB_QdS_blVoldH3TaM1WznWhs-MI5Wy1h6hYV9d1Hq2-GVuNCItx0TBqkstA=s1600
*CDS Gene Sequence (paste as text only):
ATGGACGTTGGTATCGTTGGTCTGGGTGTTATGGGTGCGAACCTGGCGCTGAATATCGCG
GAAAAGGGCTTCCACGTTGCTGTTTTCAACCGTACGTATTCTAAGGCGGAATCTTTCATC
AAGGAACACGCGTCTGCACCGTTCGCCGCTAATCTCAAAGCGTACGAAACCATGAAATCT
TTTATCATGGCGATTAAGAAGCCGCGTAAAATCTTCATCCTGGTTCAAGCGGGTGCGGCG
ACCGACGCGACCATTGAACAGCTGAAAGGTGTTCTGGAAAAAGACGACATCATCGTTGACACCGGTAACGCCCATTTCAAGGACCAAACCCGTCGTGCGGAGCAGCTGGAAGCAGCGGGCCTCCGTTTCCTGGGCATGGGTATCAGCGGTGGCGAAGAAGGTGCCCGTAAAGGCCCAGCGTTCTTCCCAGGTGGTACCCTGTCTGTTTGGAAAGAGATCCAGCCGGTTGTTGAAGCTGCGGCAGCTAAAGCGGATGACGGTCGTCCATGTGTTACTATGAACGGTCGCGGTGGTGCAGGCAGCTGTGTTAAAATGTACCACAATGCAGGCGAATACGCGGTTCTGCAGGTTTGGGGCGAAGCGTTCGACGTTCTCCGCGCGATGGGTCTCTCTAATGACGAGGTTGCGGCGGTTTTCGAGGACTGGAAAGCGAAAGGTTTCCTCACCTCTTACATGCTGGATATCTCTATTGTTGCCGCTCGTGCGAAAGTTGCGGACGGTTCCCACCTCTCTGAGCACGTTAAGGACTGCATCGGCTCTAAAGGTACGGGCCTGTGGTCCGCACAGGAGGCGCTCGAAGTGGGCGTGCCAGCGCCGAGCCTGAACATGGCGGTCCTGTCTCGTCAGATGACCATGTACAAGTCTGAACGTGAACTGAACGCAAAGACTCTGCCGTTCGTTCTGAAAGTTCCAGGTTACGCGATCAAAGACAAATCTCCGAATGCGCCGGAGATCCGTCAGCTCTACCACGCTGTTTCTATCGCAATCATCGCGTGCTACGCGCAAATGTTCCAGTGCCTGCGTGCACTCGATAAAGTTTACGACTTTGGCCTGGACCTCCCTGCAACGATCGCGACCTTCCGTGCGGGTTGCATCCTGAAGGGTTACCTGCTCCAACCAATGACCGAGGCCTTTGCGAAAAATGTTAAACTGTCTAATCTGCTGTGTGCGTTTGAAAAGGAAATCCGCGAAGGTCTCCAGAGCTATCGTGATATCCTGGGTTTCATTACCTCTAAGACTGCACTGACGCTGCCAGTTCTGTCCGCCTCTCTCGTTTACGTTAATGGTATGTTTACCCCGACCCTGCGCTATGGTCAGCTGGTTTCTCTGCAGCGTGACGTTTTTGGTCGTCATGGCTATGAGCGTCTGGACAAGGAAGGCCGTGAGTCTCACCAGTGGCCGGAGCTGCAGTAA
*GC% Content for gene:
54%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
ATGGACGTTGGTATCGTTGGTCTGGGTGTTATGGGTGCGAACCTGGCGCTGAATATCGCG
GAAAAGGGCTTCCACGTTGCTGTTTTCAACCGTACGTATTCTAAGGCGGAATCTTTCATC
AAGGAACACGCGTCTGCACCGTTCGCCGCTAATCTCAAAGCGTACGAAACCATGAAATCT
TTTATCATGGCGATTAAGAAGCCGCGTAAAATCTTCATCCTGGTTCAAGCGGGTGCGGCG
ACCGACGCGACCATTGAACAGCTGAAAGGTGTTCTGGAAAAAGACGACATCATCGTTGACACCGGTAACGCCCATTTCAAGGACCAAACCCGTCGTGCGGAGCAGCTGGAAGCAGCGGGCCTCCGTTTCCTGGGCATGGGTATCAGCGGTGGCGAAGAAGGTGCCCGTAAAGGCCCAGCGTTCTTCCCAGGTGGTACCCTGTCTGTTTGGAAAGAGATCCAGCCGGTTGTTGAAGCTGCGGCAGCTAAAGCGGATGACGGTCGTCCATGTGTTACTATGAACGGTCGCGGTGGTGCAGGCAGCTGTGTTAAAATGTACCACAATGCAGGCGAATACGCGGTTCTGCAGGTTTGGGGCGAAGCGTTCGACGTTCTCCGCGCGATGGGTCTCTCTAATGACGAGGTTGCGGCGGTTTTCGAGGACTGGAAAGCGAAAGGTTTCCTCACCTCTTACATGCTGGATATCTCTATTGTTGCCGCTCGTGCGAAAGTTGCGGACGGTTCCCACCTCTCTGAGCACGTTAAGGACTGCATCGGCTCTAAAGGTACGGGCCTGTGGTCCGCACAGGAGGCGCTCGAAGTGGGCGTGCCAGCGCCGAGCCTGAACATGGCGGTCCTGTCTCGTCAGATGACCATGTACAAGTCTGAACGTGAACTGAACGCAAAGACTCTGCCGTTCGTTCTGAAAGTTCCAGGTTACGCGATCAAAGACAAATCTCCGAATGCGCCGGAGATCCGTCAGCTCTACCACGCTGTTTCTATCGCAATCATCGCGTGCTACGCGCAAATGTTCCAGTGCCTGCGTGCACTCGATAAAGTTTACGACTTTGGCCTGGACCTCCCTGCAACGATCGCGACCTTCCGTGCGGGTTGCATCCTGAAGGGTTACCTGCTCCAACCAATGACCGAGGCCTTTGCGAAAAATGTTAAACTGTCTAATCTGCTGTGTGCGTTTGAAAAGGAAATCCGCGAAGGTCTCCAGAGCTATCGTGATATCCTGGGTTTCATTACCTCTAAGACTGCACTGACGCTGCCAGTTCTGTCCGCCTCTCTCGTTTACGTTAATGGTATGTTTACCCCGACCCTGCGCTATGGTCAGCTGGTTTCTCTGCAGCGTGACGTTTTTGGTCGTCATGGCTATGAGCGTCTGGACAAGGAAGGCCGTGAGTCTCACCAGTGGCCGGAGCTGCAGTAA
*GC% Content for gene (codon optimized): 54%

Do Not Need this info for Spring (but still copy these lines to your Target page for now)
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)
-- 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 'tail' primers (this is just 2 sequences):
**