TargetF15-+6-phosphogluconate+dehydrogenase,+decarboxylating,+putative+(Leishmania+major)


 * *Target (protein/gene name): ** 6-phosphogluconate dehydrogenase


 * *NCBI Gene # or RefSeq#: **12980488


 * *Protein ID (NP or XP #) or Wolbachia#: ** [|XP_003722728.1]


 * *Organism (including strain): ** // Leishmania major //


 * *Etiologic Risk Group: ** Risk Group 2

// Leishmania major // is a protozoan parasite, which is responsible for Leishmaniasis, a cutaneous and visceral intracellular protozoal infection. Cutaneous infections include skin rashes and sores, while visceral infections include splenomegaly and gastromegaly along with anemia. An estimated 1.3 million new cases and 20,000 to 30,000 deaths occur annually. The pathogens exist in two parasitic stages. The promastigote stage is characterized by the infection of the host by the bite of a sand-fly. Through the flagella, glycoproteins, and receptors on its membrane, the pathogen in the promastigote stage allows for the attraction of macrophages. The amastigote stage appears after the promastigote is incorporated into a macrophage. During the amastigote stage, the pathogen is present through an organism that multiplies through binary fission and spreads.
 * * **** Disease Information: **

http://tdrtargets.org/targets/view?gene_id=23670
 * *Link to TDR Targets page **

http://www.ncbi.nlm.nih.gov/gene/12980488 http://www.uniprot.org/uniprot/Q9NGR0
 * *Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): **

Substantial research has proven 6-phospho-gluconate dehydrogenase to play a profound role in the pentose phosphate pathway, glutathione metabolism, carbon metabolism, and the biosynthesis of secondary metabolites. In vivo experiments have also concluded the inhibition of 6-phosphogluconate dehydrogenase to significantly reduce fitness in bloodstream forms in other organisms.
 * *Essentiality of this protein: **
 * *Is it a monomer or multimer as biological unit? (make prediction at ** [] ): Dimer


 * *Complex of proteins **** ?: ** Dimer


 * *Druggable Target **** (list number or cite evidence from a paper/database showing druggable in another organism): ** 0.8 (TDR targets link)


 * *EC#: ** 1.1.1.44

http://www.brenda-enzymes.info/enzyme.php?ecno=1.1.1.44
 * *Link to BRENDA EC# page: **

http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/6phosphoglucdehydro74.pdf http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Bulletin/1/mak038bul.pdf http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/6phosphoglucdehydro74.pdf http://www.sigmaaldrich.com/catalog/product/sigma/mak038?lang=en®ion=US
 * -- ****Show screenshot of BRENDA enzyme mechanism schematic **
 * *Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **Spectrophotometric Assay
 * -- link to Sigma/company page for assay: **
 * -- links to assay reagents (substrates) pages: **
 * --- List cost and quantity of substrate reagents, supplier, and catalog # **

-- PDB for Homology model: 1PGJ -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB: Query Coverage: 96% coverage Max % Identities: 73% identity % Positives: 83% positive identity  Chain used for homology: A
 * Structure **** (PDB or Homology model) **


 * Homology Model IMC:**

 Model: Clash score= 11.64, Molprobity score = 2.69
Acetyl-CoA, High concentrations of palmitoyl-CoA, ATP CoA, 1-Fluoro-2,4-dinitrobenzene (irreversible), Pyridoxal 5’-phosphate with respect to ZF (competitively) and NAD or NADP (noncompetitive), L. mesenteroides G6PD, unlike mammalian G6PDs, is unaffected by steroids.
 * Current Inhibitors **** : **

http://www.ncbi.nlm.nih.gov/pubmed/14698432
 * Expression Information **** (has it been expressed in bacterial cells): ** Has been expressed in E. Coli cells


 * Purification Method **** : ** Nickel Column Chromatography

1 MSNDLGIIGLGVMGANLALNIAEKGFKVAVFNRTYAKTTSFLKEHESEKFAANLNGYETM 61 KEFAASLKKPRRAFILVQAGAATDSTIEQLKEVFENGDIIIDTGNANFKDQDKRAAQLES 121 QGLRFLGMGISGGEEGARKGPAFFPGGTPSVWEEVRPIVEAAAAKAEDGRPCVTFNGKGG 181 AGSCVKMYHNAGEYAVLQIWGEAYSALLAFGFDNDQIADVFESWKADGFLKSYMLDISIA 241 ACRAREATGNYLSEKVKDRIGSKGTGLWSAQEALEIGVPAPSLNMAVISRQMTMYKGERI 301 ANCKAFPNFPRGPSEEATDKSPNSPEAKKLYHAVSLCIIASYAQMFQCLRELDKVYGFGL 361 NLPATIATFRAGCILQGYLLGPMTKAFEENPNLPNLMDAFTKEIAAGLDDCRQILAKLTV 421 NTAVSLPVMMASLSYINAMYTETLPYGQLVSLQRDVFGRHGYERTDKDGRESFEWPALQX
 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence: **


 * *Length of your protein in Amino Acids: ** 480 aa


 * Molecular Weight of your protein in kiloDaltons using the ** [|**Expasy ProtParam**] ** website: ** 52314.8 kDa

code Ext. coefficient   51840 Abs 0.1% (=1 g/l)  0.991, assuming all pairs of Cys residues form cystines
 * Molar Extinction coefficient of your protein at 280 nm wavelength: **

Ext. coefficient   51340 Abs 0.1% (=1 g/l)  0.981, assuming all Cys residues are reduced code

code ATGTCGAACGACCTCGGCATTATCGGTCTCGGCGTCATGGGCGCGAATCTCGCCCTGAACATCGCCGAGA AGGGATTTAAAGTTGCCGTCTTCAACCGCACCTACGCGAAGACGACGTCGTTTCTCAAGGAGCATGAGAG CGAGAAATTTGCCGCCAACCTGAATGGATACGAGACCATGAAGGAGTTCGCTGCGTCCCTCAAGAAGCCG CGCCGCGCGTTCATTCTCGTCCAGGCCGGCGCCGCTACTGACTCTACAATCGAGCAGCTCAAGGAAGTGT TCGAGAACGGCGACATCATAATCGACACTGGTAATGCGAACTTCAAGGACCAGGACAAGCGCGCGGCTCA GTTGGAGAGCCAGGGTCTCCGCTTCCTCGGCATGGGCATCTCTGGTGGTGAGGAGGGTGCGCGCAAGGGG CCGGCCTTCTTCCCGGGTGGCACACCAAGCGTGTGGGAGGAGGTGCGCCCGATTGTGGAGGCGGCTGCGG CCAAGGCTGAGGACGGACGCCCGTGCGTGACTTTCAACGGCAAGGGCGGCGCTGGCTCCTGCGTGAAGAT GTATCACAACGCTGGGGAGTACGCCGTGCTGCAGATCTGGGGTGAGGCGTACAGCGCCCTGCTTGCCTTC GGCTTCGACAACGATCAGATCGCCGACGTGTTCGAGTCGTGGAAGGCGGATGGCTTCCTTAAATCCTACA TGCTCGACATCTCCATTGCCGCTTGCCGAGCGAGGGAGGCAACAGGCAACTATTTGTCAGAGAAGGTCAA GGACCGCATCGGCTCCAAGGGCACCGGCCTCTGGTCTGCCCAGGAGGCTCTGGAGATTGGTGTGCCAGCG CCCTCGCTCAACATGGCCGTCATCTCACGCCAGATGACCATGTACAAAGGGGAGCGCATCGCCAACTGCA AGGCGTTTCCTAACTTCCCTCGTGGCCCTTCTGAGGAAGCCACGGACAAGTCCCCGAACTCCCCCGAAGC GAAGAAGCTGTACCACGCCGTCAGCCTTTGTATCATCGCGAGCTACGCGCAGATGTTCCAGTGCTTGCGC GAGCTGGACAAGGTGTATGGATTCGGCCTGAACCTGCCCGCCACGATCGCCACCTTCCGCGCCGGCTGCA TTCTGCAGGGCTACCTGCTAGGGCCTATGACCAAGGCCTTCGAGGAGAACCCGAACCTGCCCAACCTGAT GGACGCCTTCACCAAGGAGATAGCGGCGGGCCTTGATGACTGCCGTCAGATTCTCGCCAAGCTCACAGTG AACACGGCAGTGTCACTGCCGGTCATGATGGCCTCACTGTCCTACATCAACGCCATGTACACGGAGACTC TTCCGTACGGACAGCTGGTGTCGTTGCAGCGCGACGTCTTTGGCCGCCACGGCTACGAGCGCACGGACAA GGACGGTCGTGAGTCGTTCGAATGGCCCGCACTGCAGTAG code
 * TMpred graph Image ** ( [] ). Input your amino acid sequence to it.
 * * **** CDS Gene Sequence: **
 * * **** GC% Content for gene (codon optimized): ** 60.4 %

1 ATGTCTAACGACCTGGGTATCATTGGCCTGGGTGTTATGGGCGCGAACCTGGCGCTGAAC 61 ATCGCGGAAAAGGGCTTCAAGGTTGCGGTTTTTAACCGTACCTATGCGAAAACCACCAGC 121 TTCCTCAAAGAACACGAATCTGAAAAATTCGCTGCCAACCTGAACGGTTACGAAACCATG 181 AAGGAATTCGCGGCCTCCCTCAAAAAGCCGCGTCGTGCGTTCATCCTGGTCCAGGCGGGT 241 GCCGCGACGGACAGCACTATCGAACAGCTGAAAGAAGTATTCGAGAATGGTGACATCATC 301 ATCGACACTGGTAACGCGAACTTTAAGGACCAAGACAAACGTGCGGCACAGCTGGAATCT 361 CAGGGTCTGCGTTTCCTCGGTATGGGCATCTCTGGTGGCGAAGAAGGTGCGCGTAAAGGT 421 CCGGCGTTCTTTCCTGGCGGTACCCCGTCTGTCTGGGAAGAAGTCCGCCCGATCGTTGAA 481 GCAGCCGCAGCAAAAGCGGAAGACGGTCGTCCTTGCGTAACCTTCAATGGCAAAGGTGGT 541 GCGGGCTCTTGCGTTAAAATGTATCACAACGCGGGTGAATACGCGGTTCTGCAAATCTGG 601 GGTGAAGCGTACTCTGCGCTGCTGGCGTTCGGTTTCGACAACGACCAGATTGCGGACGTT 661 TTCGAGTCCTGGAAAGCCGACGGTTTCCTGAAGTCCTACATGCTGGACATCTCTATTGCG 721 GCGTGCCGTGCGCGCGAAGCAACCGGCAATTACCTGTCTGAGAAAGTGAAAGACCGTATC 781 GGTTCTAAAGGTACCGGTCTGTGGTCTGCACAGGAAGCGCTGGAAATCGGTGTTCCGGCA 841 CCGTCTCTGAATATGGCGGTTATCTCTCGTCAAATGACCATGTACAAGGGTGAGCGTATC 901 GCCAACTGCAAAGCGTTCCCGAACTTCCCGCGTGGTCCGTCTGAAGAAGCGACCGATAAA 961 TCTCCGAATTCTCCGGAAGCGAAAAAACTGTACCACGCCGTATCTCTGTGCATCATTGCG 1021 TCCTACGCCCAGATGTTCCAGTGCCTGCGTGAACTGGACAAAGTTTACGGCTTCGGTCTG 1081 AACCTGCCGGCGACCATCGCGACGTTCCGTGCGGGTTGTATCCTCCAGGGCTACCTGCTG 1141 GGTCCGATGACCAAAGCCTTTGAAGAAAATCCGAACCTCCCTAACCTGATGGACGCGTTC 1201 ACCAAAGAAATCGCGGCTGGTCTGGACGACTGTCGTCAGATCCTCGCGAAACTCACCGTT 1261 AACACGGCGGTTTCTCTCCCGGTCATGATGGCCTCTCTGAGCTACATCAACGCTATGTAT 1321 ACTGAAACCCTCCCATACGGCCAGCTCGTCTCTCTGCAGCGTGACGTATTTGGTCGTCAC 1381 GGTTATGAACGTACCGACAAAGATGGCCGTGAATCTTTCGAATGGCCGGCTCTGCAATAA
 * *CDS Gene Sequence (codon optimized): **


 * * **** GC% Content for gene (codon optimized): ** 54.2 %

1 ATGTCTAACGACCTGGGTATC 21 2 GTTCAGCGCCAGGTTCGCGCCCATAACACCCAGGCCAATGATACCCAGGTCGTTAGACAT 60 3 CGAACCTGGCGCTGAACATCGCGGAAAAGGGCTTCAAGGTTGCGGTTTTTAACCGTACCT 60 4 CAGATTCGTGTTCTTTGAGGAAGCTGGTGGTTTTCGCATAGGTACGGTTAAAAACCGCAA 60 5 CTTCCTCAAAGAACACGAATCTGAAAAATTCGCTGCCAACCTGAACGGTTACGAAACCAT 60 6 GCACGACGCGGCTTTTTGAGGGAGGCCGCGAATTCCTTCATGGTTTCGTAACCGTTCAGG 60 7 AAAAAGCCGCGTCGTGCGTTCATCCTGGTCCAGGCGGGTGCCGCGACGGACAGCACTATC 60 8 GATGATGATGTCACCATTCTCGAATACTTCTTTCAGCTGTTCGATAGTGCTGTCCGTCGC 60 9 CGAGAATGGTGACATCATCATCGACACTGGTAACGCGAACTTTAAGGACCAAGACAAACG 60 10 GAGGAAACGCAGACCCTGAGATTCCAGCTGTGCCGCACGTTTGTCTTGGTCCTTAAAGTT 60 11 CAGGGTCTGCGTTTCCTCGGTATGGGCATCTCTGGTGGCGAAGAAGGTGCGCGTAAAGGT 60 12 TTCTTCCCAGACAGACGGGGTACCGCCAGGAAAGAACGCCGGACCTTTACGCGCACCTTC 60 13 CCCGTCTGTCTGGGAAGAAGTCCGCCCGATCGTTGAAGCAGCCGCAGCAAAAGCGGAAGA 60 14 CCCGCACCACCTTTGCCATTGAAGGTTACGCAAGGACGACCGTCTTCCGCTTTTGCTGCG 60 15 GCAAAGGTGGTGCGGGCTCTTGCGTTAAAATGTATCACAACGCGGGTGAATACGCGGTTC 60 16 CGAACGCCAGCAGCGCAGAGTACGCTTCACCCCAGATTTGCAGAACCGCGTATTCACCCG 60 17 CGCTGCTGGCGTTCGGTTTCGACAACGACCAGATTGCGGACGTTTTCGAGTCCTGGAAAG 60 18 TAGAGATGTCCAGCATGTAGGACTTCAGGAAACCGTCGGCTTTCCAGGACTCGAAAACGT 60 19 TCCTACATGCTGGACATCTCTATTGCGGCGTGCCGTGCGCGCGAAGCAACCGGCAATTAC 60 20 GTACCTTTAGAACCGATACGGTCTTTCACTTTCTCAGACAGGTAATTGCCGGTTGCTTCG 60 21 ACCGTATCGGTTCTAAAGGTACCGGTCTGTGGTCTGCACAGGAAGCGCTGGAAATCGGTG 60 22 TTTGACGAGAGATAACCGCCATATTCAGAGACGGTGCCGGAACACCGATTTCCAGCGCTT 60 23 GGCGGTTATCTCTCGTCAAATGACCATGTACAAGGGTGAGCGTATCGCCAACTGCAAAGC 60 24 CGGTCGCTTCTTCAGACGGACCACGCGGGAAGTTCGGGAACGCTTTGCAGTTGGCGATAC 60 25 CGTCTGAAGAAGCGACCGATAAATCTCCGAATTCTCCGGAAGCGAAAAAACTGTACCACG 60 26 ACATCTGGGCGTAGGACGCAATGATGCACAGAGATACGGCGTGGTACAGTTTTTTCGCTT 60 27 CGTCCTACGCCCAGATGTTCCAGTGCCTGCGTGAACTGGACAAAGTTTACGGCTTCGGTC 60 28 CAACCCGCACGGAACGTCGCGATGGTCGCCGGCAGGTTCAGACCGAAGCCGTAAACTTTG 60 29 CGTTCCGTGCGGGTTGTATCCTCCAGGGCTACCTGCTGGGTCCGATGACCAAAGCCTTTG 60 30 GTGAACGCGTCCATCAGGTTAGGGAGGTTCGGATTTTCTTCAAAGGCTTTGGTCATCGGA 60 31 CCTGATGGACGCGTTCACCAAAGAAATCGCGGCTGGTCTGGACGACTGTCGTCAGATCCT 60 32 TGACCGGGAGAGAAACCGCCGTGTTAACGGTGAGTTTCGCGAGGATCTGACGACAGTCGT 60 33 CGGTTTCTCTCCCGGTCATGATGGCCTCTCTGAGCTACATCAACGCTATGTATACTGAAA 60 34 ACGCTGCAGAGAGACGAGCTGGCCGTATGGGAGGGTTTCAGTATACATAGCGTTGATGTA 60 35 TCGTCTCTCTGCAGCGTGACGTATTTGGTCGTCACGGTTATGAACGTACCGACAAAGATG 60 36 TTATTGCAGAGCCGGCCATTCGAAAGATTCACGGCCATCTTTGTCGGTACGTTCATAA 58
 * Primer design results for pNIC-Bsa4 cloning: **

Forward: 5' TACTTCCAATCCATGTCTAACGACCTGGGTATCATTGGC 3' Reverse: 5' TATCCACCTTTACTGTTATTGCAGAGCCGGCCAT 3'
 * Primer design results for 'tail' primers (this is just 2 sequences): **