TargetF15-+S-adeosylmethionine+syntheses(M.+tuberculosis)

***Target (protein/gene name)**: S-adenosylmethionine synthetase

***NCBI Gene # or RefSeq#**: Tuberculist / Rv1392

***Protein ID (NP or XP #) or Wolbachia#**: ***Organism (including strain):** //M. tuberculosis //

Tuberculosis (TB) is a contagious- infectious disease that is either active or dormant depending on the individual’s ability to defend itself against the pathogen. This causes those in developing countries, young/aging populations and immunocompromised to be most at risk [1]. Active TB takes two additional forms- pulmonary and extrapulmonary (outside the lungs), with extrapulmonary being noninfectious. An individual infected with active pulmonary tuberculosis is able to spread the pathogen through the air by sneezing, coughing or talking. Repeated contact may be necessary for infection, but only about 5% of infected individuals actually develop TB [2]. Despite this roughly a third of the world’s population is estimated to be infected with //Mycobacterium tuberculosis//, the pathogen responsible for TB, leading to approximately 9 million new cases and 3 million deaths per year [1].
 * */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=6457 __

[|Link to Gene Database page (NCBI], EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **Essentiality of this protein:** Essential Reference: Comparative protein modeling of methionine S-adenosyltransferase (MAT) enzyme from Mycobacterium tuberculosis: a potential target for antituberculosis drug discovery

**Complex of proteins?**: 0.5 ***EC#**: <span style="background-color: #ffffff; font-family: arial,sans,sans-serif; font-size: 11.7px;">2.5.1.6 (methionine adenosyltransferase) same-same
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">Is it a monomer or multimer as biological unit? (make prediction at @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): **
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **


 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif; line-height: 1.5;">Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **

<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">[|http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3836674/#R7]
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">-- link to Sigma (or other company) page for assay (see Sigma links below) **
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">-- -or link (or citation) to paper that contains assay information: **
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">-- links to assay reagents (substrates) pages. **
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">--- List cost and quantity of substrate reagents, supplier, and catalog # **


 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">Structure (PDB or Homology model) **
 * <span style="font-family: Arial,Helvetica,sans-serif; line-height: 1.5;">Current Inhibitors **<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif; line-height: 1.5;">:

<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">**Expression Information (has it been expressed in bacterial cells)**: Yes, has crystal structure

<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">**Purification Method**: <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">***Amino Acid Sequence (paste as text only - not as screenshot or as 'code')**:
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">Image of protein (PyMol with features delineated and shown separately): **

code GPGSMVSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVTTSAKEAFADITNTVR ARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARVEGAADPLDSQGAGDQGLMFGYAINATPELMPLPIA LAHRLSRRLTEVRKNGVLPYLRPDGKTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHET LDASTVRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMRWVAKNVVAAGLAERV EVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGAIIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDK VDDLKRAI code

<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">***length of your protein in Amino Acids:** <span style="background-color: #ffffff; font-family: arial,sans,sans-serif;">261 aa

<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">**Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website**: code 43444.1 code

<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">**Molar Extinction coefficient of your protein at 280 nm wavelength**: code Ext. coefficient   30035 Abs 0.1% (=1 g/l)  0.691, assuming all pairs of Cys residues form cystines

Ext. coefficient   29910 Abs 0.1% (=1 g/l)  0.688, assuming all Cys residues are reduced code


 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">TMpred graph Image (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. ****<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif; line-height: 1.5;">*CDS Gene Sequence (paste as text only): **

<span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">**CDS Gene:**
<span style="background-color: #00ff00; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">ATG <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">__GTTTCTGAAAAAGGTCGT__CTGTTTACGTCTGAGTCTGTTACGGAAGGCCATCCTGACAAAATTTGCGACG <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">CTATCTCTGACTCTGTACTGGATGCACTCCTGGCGGCAGACCCTCGTTCTCGTGTTGCGGTTGAGACTCTCGT <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">AACCACCGGTCAGGTTCACGTTGTTGGTGAAGTTACCACCTCTGCAAAAGAAGCGTTTGCGGATATCACTAAC <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">ACGGTCCGTGCGCGTATTCTGGAAATTGGCTACGACTCTTCTGACAAGGGTTTCGATGGTGCGACGTGCGGC <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">GTGAACATCGGTATCGGTGCGCAGTCTCCTGATATTGCGCAGGGTGTTGACACCGCACACGAAGCGCGTGTG <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">GAAGGTGCTGCAGATCCTCTGGACTCTCAGGGTGCCGGTGACCAGGGCCTGATGTTCGGTTACGCAATCAAT <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">GCGACCCCTGAACTGATGCCGCTCCCGATCGCGCTGGCCCATCGCCTGTCTCGTCGTCTCACTGAGGTTC <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">GTAAAAACGGTGTTCTCCCGTACCTCCGCCCAGATGGTAAGACCCAGGTTACTATCGCGTACGAGGATAACGT <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">TCCGGTTCGTCTCGACACTGTGGTAATCTCTACCCAGCACGCGGCTGACATCGACCTGGAGAAAACCCTCGA <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">TCCGGACATCCGCGAGAAAGTTCTCAACACTGTCCTGGATGACCTCGCACATGAGACGCTGGACGCATCCAC <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">CGTTCGTGTTCTGGTAAATCCTACGGGCAAATTCGTTCTGGGTGGTCCGATGGGTGATGCGGGTCTGACTGG <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">TCGCAAGATCATCGTAGACACCTACGGCGGTTGGGCCCGTCACGGTGGTGGTGCGTTCAGCGGCAAAGACC <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">CGTCTAAGGTTGACCGTTCTGCGGCCTATGCCATGCGTTGGGTTGCGAAAAACGTCGTGGCAGCAGGTCTGG <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">CGGAGCGTGTAGAAGTTCAAGTCGCGTATGCAATCGGTAAAGCGGCACCTGTTGGTCTGTTCGTTGAAACTTT <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">CGGCACCGAAACGGAAGATCCGGTAAAAATCGAAAAAGCGATTGGTGAGGTTTTTGATCTGCGTCCGGGTGC <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">CATCATTCGTGATCTGAATCTGCTCCGTCCAATCTACGCGCCGACCGCAGCGTACGGCCACTTTGGTCGTACC <span style="background-color: #ffffff; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">GACGTTGAACTGCCGTGGGAACAACTGGACAAGGTCGACG__ATCTCAAACGTGCGATC__ <span style="background-color: #ff0000; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">__TAA__ <span style="background-color: #ff0000; font-family: Arial; font-size: 18.6667px; vertical-align: baseline;">

KTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDAST
VRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMR WVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGA IIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI

Methionine added to beginning of sequence for E coli MVSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVT TSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARV EGAADPLDSQGAGDQGLMFGYAINATPELMPLPIALAHRLSRRLTEVRKNGVLPYLRPDG KTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDAST VRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMR WVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGA IIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI
 * <span style="background-color: #ffffff; font-family: Arial,Helvetica,sans-serif;">Primer design results for 'tail' primers (this is just 2 sequences): **

Forward Primer: 5’ TACTTCCAATCC <span style="background-color: #ffffff; color: #ff0000; font-family: Arial; font-size: 13.3333px; vertical-align: baseline;">ATGGTTTCTGAAAAAGGTCG 3’ __32__ bp GC Content __40.6__% 0 mM Mg 2+ T m __60.3__ o C1.5 mM Mg 2+ T m __68.1__ o C 2 mM Mg 2+ T m __68.5__ o C 4 mM Mg 2+ T m __69.4__ o C 6 mM Mg 2+ T m __69.9__ o C

__Reverse Primer__: 5’ TATCCACCTTTACTG TTAGATCGCACGTTTGAGAT 3’ __32__ bp GC Content __40__% 0 mM Mg 2+ T m __60.7__ o C1.5 mM Mg 2+ T m __68.6__ o C 2 mM Mg 2+ T m __69__ o C 4 mM Mg 2+ T m __69.8__ o C 6 mM Mg 2+ T m __70.3__ o C

<span style="background-color: #ffffff; font-family: arial,sans,sans-serif;">References:

<span style="font-family: Calibri,sans-serif; font-size: 10pt;">[1] Ducati, R. G.; Ruffino-Netto, A.; Basso, L. A.; Santos, D. S., The resumption of consumption -- a review on tuberculosis. //Mem Inst Oswaldo Cruz// **2006,**//101// (7), 697-714.