Target+- Glyceraldehyde-3-phosphate+dehydrogenase+(T.+Cruzi)

Glyceraldehyde-3-phosphate dehydrogenase Tc00.1047053503687.20 3535978 XP_806067 (or XP_806067.1) //Trypanosoma cruzi// Risk Group 2 (RG2) - Parasitic Agents Chagas disease is a parasitic disease caused by the organism //Trypanosoma cruzi// that is prevalent throughout Central and South America. It is estimated that Chagas disease is affecting 8 to 11 million people and it is predominant in rural and impoverished areas. Recently though, it has been spreading out from Latin America into North America, as evidenced by the increased risk for Chagas disease in south Texas. It is possible that there could already be hundreds of people infected in Texas but have yet to be diagnosed due to the difficulty in symptom recognition. Chagas disease has an acute phase and a chronic phase. In the acute phase, harsh symptoms are uncommon and often people are symptom free or experience only flu-like symptoms. Some symptoms during this phase can include “fever, fatigue, body aches, headache, rash, loss of appetite, diarrhea, and vomiting.” The chronic phase can remain silent for months to a lifetime depending on the specific case but people can develop two main kinds of complications: cardiac complications and intestinal complications. Treatment in the acute phase usually consists of antiparasitic treatment with the drug Benznidazole or its alternative Nifurtimox. Often when patients lapse into the chronic phase, symptomatic treatment is also recommended to deal with the cardiac or intestinal problems that can arise. Chagas disease is transmitted primarily by triatomine bugs, a blood-sucking insect vector that is often given the name “kissing bugs” because of their tendencies to feed on the faces of sleeping people. The bugs become infected themselves by biting an infected person or animal and then pass it on through their feces, which enters the body through cuts or mucous membranes. It can also be transmitted through blood transfusions, congenital fluids, organ transplants, and the consumption of food contaminated with the feces of triatomine bugs that have the disease. The population that is most susceptible is the geographic region where the triatomine bugs make their home due to their pivotal role as vectors. Once in the body, the //T. cruzi// parasite, known at this point as a metacyclic trypomastigotes, transform into amastigotes. The amastigotes then replicate by binary fission in the infected cells. They then transform into trypomastigotes and burst into the bloodstream where they can then infect other cells and repeat the previous process of replication. If a triatomine bug then bites a person, the bug will become infected and can pass it on to the next victim. In Central and South America, methods have been put into place to try to control the spread of Chagas disease. In an attempt to limit the amount of people exposed to the triatomine bugs, housing conditions have been improved and many have taken to spraying insecticide in hopes of killing infected bugs. Also, early detection protocols and more stringent blood donation screenings are being put into place.
 * Target (protein/gene name): **
 * NCBI Gene # or RefSeq#: **
 * Gene ID:**
 * Protein ID (NP or XP #) or Wolbachia#: **
 * Organism: **
 * Etiologic Risk Group (see link below): **
 * Background/Disease Information: **

significant loss of fitness in bloodstream forms (3 days) from a study of an African trypanosome [alsford] significant loss of fitness in bloodstream forms (6 days) from a study of an African trypanosome [alsford] significant gain of fitness in procyclic forms from a study of an African trypanosome [alsford]
 * Essentiality of this protein: **


 * Complex of enzymes: **

1.2.1.12 []
 * EC#: **
 * Link to BRENDA EC# page: **

The enzyme assay that will be performed will be spectrophotometric.
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **

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 * -- link to Sigma (or other company) page for assay or assay reagents (substrates) **
 * -- Reagent Costs**
 * **Reagent** || **Sigma Product Number** || **Amount** || **Cost** ||
 * [|Triethanolamine hydrochloride] || T1502 || 100g || $29.80 ||
 * [|D-(−)-3-Phosphoglyceric acid disodium salt] || P8877 || 10mg || $13.30 ||
 * [|L-Cysteine hydrochloride monohydrate] || C7880 || 500mg || $8.70 ||
 * [|Sodium bicarbonate] || S8875 || 500g || $31.00 ||
 * [|Magnesium sulfate heptahydrate] || M1880 || 500g || $36.00 ||
 * [|β-Nicotinamide adenine dinucleotide, reduced disodium salt hydrate] || N1829 || 50mg || $22.60 ||
 * [|Adenosine 5′-triphosphate disodium salt hydrate] || A7699 || 1g || $63.70 ||
 * [|3-Phosphoglyceric Phosphokinase] || P7634 || 2KU || $52.60 ||

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 * -- link to paper that contains assay information **



1QXS 1K3T
 * Structure Available (PDB or Homology model) **
 * -- PDB # or closest PDB entry if using homology model: **
 * -- Query Coverage (if not direct match): **
 * -- Max Ident (if not direct match): **

0.3 The following compounds were studied in this [|paper] and determined to have inhibitory functions. (8E)-C15:1_anacardic acid, (8E,11E)-C15:2-anacardic acid, (8E,11E,14E)-C15:3-anacardic acid, 2-(dodec-1-en-1-yl)-6-hydroxybenzoic acid, 2-pentadecyl-6-hydroxybenzoic acid, guajaverin, mangiferin, tiliroside It has been expressed in bacterial cells, E. coli BL21 (DE3) using a pET-28a+) vector. It was purified using column chromatography methods with an Ni-NTA resign. The protein was tagged with a six histidine tag which facilitated this method.
 * Druggable Target (see Databases for this): **
 * Current Inhibitors: **
 * Expression Information (has it been expressed in bacterial cells): **
 * Purification Method: **
 * Image of protein (PyMol or etc): **

>gi|71407160|ref|XP_806067.1| glyceraldehyde-3-phosphate dehydrogenase [Trypanosoma cruzi strain CL Brener] MPLLLLRWLRRTVLLFCHWRRVIRMEDVVLPVCVGINGFGPIGQAVLFSSFTDPLVSVVAINDASMSIDY IAYLLRRESSLSAGDRASVLVVGEFICIQGSQKIRVSHKHDLVEIAWRDVGVQYVVECTGLSSTRERCWG HVAGGAKGVIVAGQSADAPTLIAGANDEELKSACSVVCAGSPVAVALAPLIRLLHEQYGLEECSYTAIHG MRPVELTAGRSKNPQDWRQTRVSIDNIVPYIDNGKKTMDKIFPGLVGRISGTAFQVPVKKGCAVDMLVRL SQPVSKEMLDQALKEAASGRLNGVLSYSKEDLISCDCVPNGKLCYDATGSCSLRDGEAQKLLLWFDIDGG YAKRLLSLVVLLHQMGPSDGM
 * Amino Acid Sequence: **

371aa 40.2897kD 46410 (assuming all Cys residues are reduced)
 * length of your protein in Amino Acids: **
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: **
 * Molar Extinction coefficient of your protein at 280 nm wavelength: **


 * CDS Gene Sequence: **

ATGCCGCTGCTGCTTCTGCGGTGGCTACGACGGACAGTGTTGCTTTTTTGCCATTGGAGA AGAGTGATAAGGATGGAGGATGTTGTCTTGCCGGTTTGTGTGGGCATTAATGGCTTTGGG CCAATTGGGCAGGCCGTTCTTTTTTCCTCGTTCACGGACCCTTTAGTGAGCGTGGTGGCG ATAAACGATGCTTCCATGAGTATTGACTACATTGCATACTTGCTGCGGCGCGAAAGCTCT CTTTCTGCAGGGGACAGAGCCTCAGTGTTGGTGGTTGGGGAGTTTATCTGCATCCAGGGT TCTCAGAAAATCCGTGTTTCACATAAGCATGATCTTGTGGAGATTGCATGGCGAGATGTC GGGGTGCAGTACGTGGTGGAGTGCACGGGACTAAGCTCCACGCGAGAACGCTGCTGGGGA CATGTTGCAGGAGGTGCCAAAGGCGTCATTGTTGCAGGACAGAGTGCGGACGCCCCAACT CTCATTGCTGGTGCAAATGACGAAGAATTGAAAAGTGCGTGTTCCGTTGTGTGTGCTGGC TCCCCTGTTGCAGTCGCCCTCGCTCCTCTCATTCGTTTGCTGCATGAGCAATACGGTCTA GAAGAGTGCTCCTACACCGCCATTCACGGTATGAGACCAGTGGAGCTTACAGCAGGAAGG TCTAAAAATCCGCAAGACTGGCGCCAAACAAGAGTGAGTATTGACAACATTGTTCCTTAT ATCGACAACGGGAAAAAGACTATGGATAAAATTTTCCCAGGTCTTGTGGGACGCATCTCA GGAACTGCGTTTCAGGTCCCCGTGAAAAAAGGATGTGCCGTGGACATGTTGGTCCGGTTG AGCCAGCCTGTCTCGAAGGAAATGTTGGATCAAGCTCTGAAGGAAGCTGCTTCAGGTCGA CTCAACGGAGTGCTTTCTTACTCAAAAGAGGACCTCATTAGTTGCGACTGTGTGCCAAAT GGTAAGCTGTGTTACGACGCAACAGGTTCCTGTTCTCTGCGTGATGGGGAGGCGCAAAAA CTACTGTTATGGTTTGATATTGATGGTGGTTACGCGAAGAGGCTTCTGTCATTAGTGGTG CTTTTGCATCAGATGGGTCCCAGTGACGGGATGTAG

50.99%
 * GC% Content for gene: **
 * CDS Gene Sequence (codon optimized) - after having done Primer Design: **

ATGCCTCTGCTCCTGCTGCGTTGGCTGCGTCGTACTGTACTCCTCTTTTGCCACTGGCGC CGTGTAATCCGCATGGAAGACGTAGTTCTGCCAGTATGTGTAGGTATCAACGGTTTCGGT CCTATCGGTCAGGCGGTTCTGTTTTCTAGCTTTACCGATCCTCTGGTTTCTGTAGTTGCT ATTAACGACGCCTCTATGTCTATCGACTACATCGCGTACCTGCTCCGTCGCGAATCTTCT CTGTCTGCCGGTGACCGTGCGAGCGTTCTCGTAGTTGGTGAGTTTATTTGCATCCAGGGT TCTCAGAAAATCCGTGTTTCTCACAAACACGACCTGGTTGAAATCGCGTGGCGCGACGTT GGTGTTCAGTACGTTGTTGAATGCACCGGTCTCTCCTCTACGCGTGAACGTTGCTGGGGT CACGTGGCGGGTGGTGCTAAAGGTGTTATTGTTGCGGGTCAGTCTGCGGACGCCCCGACC CTGATCGCAGGTGCGAACGACGAAGAACTGAAATCTGCATGCTCTGTTGTTTGCGCCGGT TCCCCGGTAGCGGTTGCTCTGGCCCCACTGATCCGTCTCCTCCACGAGCAGTACGGTCTG GAAGAATGTTCTTACACCGCGATTCACGGTATGCGTCCGGTTGAACTGACCGCGGGTCGT TCTAAAAACCCGCAGGACTGGCGTCAGACCCGTGTGTCCATTGATAACATCGTTCCGTAC ATCGACAACGGTAAGAAGACCATGGACAAAATCTTCCCGGGTCTCGTCGGCCGTATTTCT GGTACTGCATTCCAGGTTCCGGTTAAGAAAGGTTGTGCGGTTGACATGCTGGTTCGTCTG TCTCAGCCGGTTTCTAAAGAAATGCTGGACCAGGCGCTGAAAGAAGCGGCGTCTGGTCGT CTGAACGGCGTTCTGTCTTACTCTAAGGAAGACCTGATCTCTTGCGACTGCGTTCCGAAC GGCAAACTGTGCTACGATGCGACCGGTTCTTGTTCTCTGCGTGATGGCGAAGCGCAAAAA CTGCTGCTGTGGTTCGACATTGACGGTGGTTACGCGAAACGCCTCCTCTCCCTGGTGGTT CTGCTGCACCAAATGGGTCCGAGCGATGGTATGTAA

54.12 % 1 ATGCCTCTGCTCCTGCTGCGTTGGCTGCGTCGTAC 35 2 TTCCATGCGGATTACACGGCGCCAGTGGCAAAAGAGGAGTACAGTACGACGCAGCCAACG 60 3 CCGTGTAATCCGCATGGAAGACGTAGTTCTGCCAGTATGTGTAGGTATCAACGGTTTCGG 60 4 TCGGTAAAGCTAGAAAACAGAACCGCCTGACCGATAGGACCGAAACCGTTGATACCTACA 60 5 GTTCTGTTTTCTAGCTTTACCGATCCTCTGGTTTCTGTAGTTGCTATTAACGACGCCTCT 60 6 TTCGCGACGGAGCAGGTACGCGATGTAGTCGATAGACATAGAGGCGTCGTTAATAGCAAC 60 7 CCTGCTCCGTCGCGAATCTTCTCTGTCTGCCGGTGACCGTGCGAGCGTTCTCGTAGTTGG 60 8 AAACACGGATTTTCTGAGAACCCTGGATGCAAATAAACTCACCAACTACGAGAACGCTCG 60 9 GGTTCTCAGAAAATCCGTGTTTCTCACAAACACGACCTGGTTGAAATCGCGTGGCGCGAC 60 10 GCGTAGAGGAGAGACCGGTGCATTCAACAACGTACTGAACACCAACGTCGCGCCACGCGA 60 11 CCGGTCTCTCCTCTACGCGTGAACGTTGCTGGGGTCACGTGGCGGGTGGTGCTAAAGGTG 60 12 CGATCAGGGTCGGGGCGTCCGCAGACTGACCCGCAACAATAACACCTTTAGCACCACCCG 60 13 GCCCCGACCCTGATCGCAGGTGCGAACGACGAAGAACTGAAATCTGCATGCTCTGTTGTT 60 14 CAGTGGGGCCAGAGCAACCGCTACCGGGGAACCGGCGCAAACAACAGAGCATGCAGATTT 60 15 TTGCTCTGGCCCCACTGATCCGTCTCCTCCACGAGCAGTACGGTCTGGAAGAATGTTCTT 60 16 CGGTCAGTTCAACCGGACGCATACCGTGAATCGCGGTGTAAGAACATTCTTCCAGACCGT 60 17 GTCCGGTTGAACTGACCGCGGGTCGTTCTAAAAACCCGCAGGACTGGCGTCAGACCCGTG 60 18 CTTCTTACCGTTGTCGATGTACGGAACGATGTTATCAATGGACACACGGGTCTGACGCCA 60 19 GTACATCGACAACGGTAAGAAGACCATGGACAAAATCTTCCCGGGTCTCGTCGGCCGTAT 60 20 CGCACAACCTTTCTTAACCGGAACCTGGAATGCAGTACCAGAAATACGGCCGACGAGACC 60 21 CGGTTAAGAAAGGTTGTGCGGTTGACATGCTGGTTCGTCTGTCTCAGCCGGTTTCTAAAG 60 22 GACCAGACGCCGCTTCTTTCAGCGCCTGGTCCAGCATTTCTTTAGAAACCGGCTGAGACA 60 23 AGAAGCGGCGTCTGGTCGTCTGAACGGCGTTCTGTCTTACTCTAAGGAAGACCTGATCTC 60 24 CGTAGCACAGTTTGCCGTTCGGAACGCAGTCGCAAGAGATCAGGTCTTCCTTAGAGTAAG 60 25 ACGGCAAACTGTGCTACGATGCGACCGGTTCTTGTTCTCTGCGTGATGGCGAAGCGCAAA 60 26 CGTTTCGCGTAACCACCGTCAATGTCGAACCACAGCAGCAGTTTTTGCGCTTCGCCATCA 60 27 CGGTGGTTACGCGAAACGCCTCCTCTCCCTGGTGGTTCTGCTGCACCAAATGGGTCCGAG 60 28 TTACATACCATCGCTCGGACCCATTTGGTGC 31
 * GC% Content for gene (codon optimized): **
 * Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **

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 * Google Docs Primer Design Log File:**


 * Primer design results for 'tail' primers (this is just 2 sequences): **