6-phosphogluconate+dehydrogenase,+decarboxylating,+putative+T.+cruzi


 * *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): **

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
 * */ Disease Information (sort of like the Intro to your Mini __Research Write__ up): **


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

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


 * 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

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

Spectrophotometric → UV Quantification [|__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__]
 * *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
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * -- 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: 1PGJ -- For Homology Model option:
 * Structure (PDB or Homology model) **

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__]

MDVGIVGLGVMGANLALNIAEKGFHVAVFNRTYSKAESFIKEHASAPFAANLKAYETMKS FIMAIKKPRKIFILVQAGAATDATIEQLKGVLEKDDIIVDTGNAHFKDQTRRAEQLEAAG LRFLGMGISGGEEGARKGPAFFPGGTLSVWKEIQPVVEAAAAKADDGRPCVTMNGRGGAG SCVKMYHNAGEYAVLQVWGEAFDVLRAMGLSNDEVAAVFEDWKAKGFLTSYMLDISIVAA RAKVADGSHLSEHVKDCIGSKGTGLWSAQEALEVGVPAPSLNMAVLSRQMTMYKSERELN AKTLPFVLKVPGYAIKDKSPNAPEIRQLYHAVSIAIIACYAQMFQCLRALDKVYDFGLDL PATIATFRAGCILKGYLLQPMTEAFAKNVKLSNLLCAFEKEIREGLQSYRDILGFITSKT ALTLPVLSASLVYVNGMFTPTLRYGQLVSLQRDVFGRHGYERLDKEGRESHQWPELQ 5,2134.4 Da
 * Purification Method : ** Nickel affinity chromatography
 * 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: ** 477 aa
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: **
 * 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

ATGGACGTTGGTATCGTTGGTCTGGGTGTTATGGGTGCGAACCTGGCGCTGAATATCGCG GAAAAGGGCTTCCACGTTGCTGTTTTCAACCGTACGTATTCTAAGGCGGAATCTTTCATC AAGGAACACGCGTCTGCACCGTTCGCCGCTAATCTCAAAGCGTACGAAACCATGAAATCT TTTATCATGGCGATTAAGAAGCCGCGTAAAATCTTCATCCTGGTTCAAGCGGGTGCGGCG ACCGACGCGACCATTGAACAGCTGAAAGGTGTTCTGGAAAAAGACGACATCATCGTTGACACCGGTAACGCCCATTTCAAGGACCAAACCCGTCGTGCGGAGCAGCTGGAAGCAGCGGGCCTCCGTTTCCTGGGCATGGGTATCAGCGGTGGCGAAGAAGGTGCCCGTAAAGGCCCAGCGTTCTTCCCAGGTGGTACCCTGTCTGTTTGGAAAGAGATCCAGCCGGTTGTTGAAGCTGCGGCAGCTAAAGCGGATGACGGTCGTCCATGTGTTACTATGAACGGTCGCGGTGGTGCAGGCAGCTGTGTTAAAATGTACCACAATGCAGGCGAATACGCGGTTCTGCAGGTTTGGGGCGAAGCGTTCGACGTTCTCCGCGCGATGGGTCTCTCTAATGACGAGGTTGCGGCGGTTTTCGAGGACTGGAAAGCGAAAGGTTTCCTCACCTCTTACATGCTGGATATCTCTATTGTTGCCGCTCGTGCGAAAGTTGCGGACGGTTCCCACCTCTCTGAGCACGTTAAGGACTGCATCGGCTCTAAAGGTACGGGCCTGTGGTCCGCACAGGAGGCGCTCGAAGTGGGCGTGCCAGCGCCGAGCCTGAACATGGCGGTCCTGTCTCGTCAGATGACCATGTACAAGTCTGAACGTGAACTGAACGCAAAGACTCTGCCGTTCGTTCTGAAAGTTCCAGGTTACGCGATCAAAGACAAATCTCCGAATGCGCCGGAGATCCGTCAGCTCTACCACGCTGTTTCTATCGCAATCATCGCGTGCTACGCGCAAATGTTCCAGTGCCTGCGTGCACTCGATAAAGTTTACGACTTTGGCCTGGACCTCCCTGCAACGATCGCGACCTTCCGTGCGGGTTGCATCCTGAAGGGTTACCTGCTCCAACCAATGACCGAGGCCTTTGCGAAAAATGTTAAACTGTCTAATCTGCTGTGTGCGTTTGAAAAGGAAATCCGCGAAGGTCTCCAGAGCTATCGTGATATCCTGGGTTTCATTACCTCTAAGACTGCACTGACGCTGCCAGTTCTGTCCGCCTCTCTCGTTTACGTTAATGGTATGTTTACCCCGACCCTGCGCTATGGTCAGCTGGTTTCTCTGCAGCGTGACGTTTTTGGTCGTCATGGCTATGAGCGTCTGGACAAGGAAGGCCGTGAGTCTCACCAGTGGCCGGAGCTGCAGTAA 54% ATGGACGTTGGTATCGTTGGTCTGGGTGTTATGGGTGCGAACCTGGCGCTGAATATCGCG GAAAAGGGCTTCCACGTTGCTGTTTTCAACCGTACGTATTCTAAGGCGGAATCTTTCATC AAGGAACACGCGTCTGCACCGTTCGCCGCTAATCTCAAAGCGTACGAAACCATGAAATCT TTTATCATGGCGATTAAGAAGCCGCGTAAAATCTTCATCCTGGTTCAAGCGGGTGCGGCG ACCGACGCGACCATTGAACAGCTGAAAGGTGTTCTGGAAAAAGACGACATCATCGTTGACACCGGTAACGCCCATTTCAAGGACCAAACCCGTCGTGCGGAGCAGCTGGAAGCAGCGGGCCTCCGTTTCCTGGGCATGGGTATCAGCGGTGGCGAAGAAGGTGCCCGTAAAGGCCCAGCGTTCTTCCCAGGTGGTACCCTGTCTGTTTGGAAAGAGATCCAGCCGGTTGTTGAAGCTGCGGCAGCTAAAGCGGATGACGGTCGTCCATGTGTTACTATGAACGGTCGCGGTGGTGCAGGCAGCTGTGTTAAAATGTACCACAATGCAGGCGAATACGCGGTTCTGCAGGTTTGGGGCGAAGCGTTCGACGTTCTCCGCGCGATGGGTCTCTCTAATGACGAGGTTGCGGCGGTTTTCGAGGACTGGAAAGCGAAAGGTTTCCTCACCTCTTACATGCTGGATATCTCTATTGTTGCCGCTCGTGCGAAAGTTGCGGACGGTTCCCACCTCTCTGAGCACGTTAAGGACTGCATCGGCTCTAAAGGTACGGGCCTGTGGTCCGCACAGGAGGCGCTCGAAGTGGGCGTGCCAGCGCCGAGCCTGAACATGGCGGTCCTGTCTCGTCAGATGACCATGTACAAGTCTGAACGTGAACTGAACGCAAAGACTCTGCCGTTCGTTCTGAAAGTTCCAGGTTACGCGATCAAAGACAAATCTCCGAATGCGCCGGAGATCCGTCAGCTCTACCACGCTGTTTCTATCGCAATCATCGCGTGCTACGCGCAAATGTTCCAGTGCCTGCGTGCACTCGATAAAGTTTACGACTTTGGCCTGGACCTCCCTGCAACGATCGCGACCTTCCGTGCGGGTTGCATCCTGAAGGGTTACCTGCTCCAACCAATGACCGAGGCCTTTGCGAAAAATGTTAAACTGTCTAATCTGCTGTGTGCGTTTGAAAAGGAAATCCGCGAAGGTCTCCAGAGCTATCGTGATATCCTGGGTTTCATTACCTCTAAGACTGCACTGACGCTGCCAGTTCTGTCCGCCTCTCTCGTTTACGTTAATGGTATGTTTACCCCGACCCTGCGCTATGGTCAGCTGGTTTCTCTGCAGCGTGACGTTTTTGGTCGTCATGGCTATGAGCGTCTGGACAAGGAAGGCCGTGAGTCTCACCAGTGGCCGGAGCTGCAGTAA
 * TMpred graph Image ** ( @http://www.ch.embnet.org/software/TMPRED_form.html ). Input your amino acid sequence to it.
 * *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): ** 54%

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): **