Target+-+Inositol-1-monophosphatase+(Mycobacterium+tuberculosis)

SuhB //Mycobacterium tuberculosis //is the bacteria which causes tuberculosis, an infectious disease that mainly affects the lungs [2]. This disease is estimated to be the cause of three million death per year worldwide [1]. The bacteria is spread to individuals by an infected person’s coughs or sneezes. Those at risk include those with HIV, diabetes, cancer, or malnutrition. Many medications exist to treat tuberculosis, though many strains of the disease are resistant to these drugs. Infected individuals must take a cocktail of various types of medications for long periods of time to prevent antibiotic resistance [2]. The mycobacterial cell wall is composed of various substances based on a phosphatidyl-myo-inositol structure. These substances are extremely important in the cell wall of mycobacterial species. They also greatly contribute to the immunopathogenesis by tuberculosis [1]. Inositol-1-monophosphatase is a critical enzyme in inositol biosynthesis in Mycobacterium tuberculosis SuhB. It synthesizes free myo-inositol from inositol-1-phosphate, a key substrate for the phosphatidylinositol synthase in mycobacteria.  Not essential via Brown, et. al. 2007
 * Target: **Inositol-1-monophosphatase
 * NCBI Gene#: ** 17725819
 * Protein ID#: ** 10.1186/1472-6807-7-55
 * Organism: **Mycobacterium tuberculosis
 * Etiologic Risk Group: **RG3
 * Background/Disease Information: **
 * Essentiality of this protein: **


 * <span style="font-family: Cambria,serif; font-size: 11pt;">Complex of proteins?: **<span style="font-family: Cambria,serif; font-size: 11pt;">No, dimerizes in presence of Mg2+ to be active
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Druggable Target: **<span style="font-family: Cambria,serif; font-size: 11pt;">Yes (0.5 Druggability in TDR Targets)

<span style="font-family: Cambria,serif; font-size: 11pt;">[]
 * <span style="font-family: Cambria,serif; font-size: 11pt;">*EC#: ** <span style="font-family: Cambria,serif; font-size: 11pt;">3.1.3.25
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Link to BRENDA EC# page: **


 * <span style="font-family: Cambria,serif; font-size: 11pt;">Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **

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 * <span style="font-family: Cambria,serif; font-size: 11pt;">-- link to Sigma (or other company) page for assay or assay reagents (substrates) **


 * <span style="font-family: Cambria,serif; font-size: 11pt;">-- ****<span style="font-family: Cambria,serif; font-size: 11pt; line-height: 1.5;">Cost and quantity of substrate reagents and supplier: **<span style="background-color: white; font-family: Cambria,serif; font-size: 11pt;">40mM pNPP (Sigma): $ <span style="font-family: Cambria,serif; font-size: 11pt;">79.60/1g

<span style="font-family: Cambria,serif; font-size: 11pt;">-- PDB # 2Q74
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Structure Available from PDB **

<span style="font-family: Cambria,serif; font-size: 11pt;">Li+ (IC50: 0.9 mM) <span style="font-family: Cambria,serif; font-size: 11pt;">Zn2+ (IC50:0.5 uM) <span style="font-family: Cambria,serif; font-size: 11pt;">Mg2+ concentrations higher than 100mM <span style="font-family: Cambria,serif; font-size: 11pt;">Yes, it has been expressed in E.Coli [1]. <span style="font-family: Cambria,serif; font-size: 11pt;"> <span style="font-family: Cambria,serif; font-size: 11pt;"> 10 20 30 40 50 60 <span style="font-family: Cambria,serif; font-size: 11pt;">MTRPDNEPAR LRSVAENLAA EAAAFVRGRR AEVFGISRAG DGDGAVRAKS SPTDPVTVVD
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Current Inhibitors: **
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Expression Information: **
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Purification Method: **<span style="font-family: Cambria,serif; font-size: 11pt;">His Tag Purification
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Image of protein : **
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Amino Acid Sequence: **

<span style="font-family: Cambria,serif; font-size: 11pt;"> 70 80 90 100 110 120 <span style="font-family: Cambria,serif; font-size: 11pt;">TDTERLLRDR LAQLRPGDPI LGEEGGGPAD VTATPSDRVT WVLDPIDGTV NFVYGIPAYA

<span style="font-family: Cambria,serif; font-size: 11pt;"> 130 140 150 160 170 180 <span style="font-family: Cambria,serif; font-size: 11pt;">VSIGAQVGGI TVAGAVADVA ARTVYSAATG LGAHLTDERG RHVLRCTGVD ELSMALLGTG

<span style="font-family: Cambria,serif; font-size: 11pt;"> 190 200 210 220 230 240 <span style="font-family: Cambria,serif; font-size: 11pt;">FGYSVRCREK QAELLAHVVP LVRDVRRIGS AALDLCMVAA GRLDAYYEHG VQVWDCAAGA

<span style="font-family: Cambria,serif; font-size: 11pt;"> 250 260 270 280 290 <span style="font-family: Cambria,serif; font-size: 11pt;">LIAAEAGARV LLSTPRAGGA GLVVVAAAPG IADELLAALQ RFNGLEPIPD

[] []
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Number of Amino Acids: **<span style="font-family: Cambria,serif; font-size: 11pt;">267
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Molecular Weight: **<span style="font-family: Cambria,serif; font-size: 11pt;">28447.1


 * <span style="font-family: Cambria,serif; font-size: 11pt;">Molar Extinction coefficient of your protein at 280 nm wavelength: **<span style="font-family: Cambria,serif; font-size: 11pt; line-height: 1.5;">45950
 * <span style="font-family: Cambria,serif; font-size: 11pt;">TMpred graph Image **<span style="font-family: Cambria,serif; font-size: 11pt;"> :
 * <span style="font-family: Cambria,serif; font-size: 11pt;">*CDS Gene Sequence (paste as text only): **
 * <span style="font-family: Cambria,serif; font-size: 11pt;">*GC% Content for gene: **<span style="font-family: Cambria,serif; font-size: 11pt; line-height: 1.5;">0.4%
 * <span style="font-family: Cambria,serif; font-size: 11pt;">*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): **
 * <span style="font-family: Cambria,serif; font-size: 11pt;">*GC% Content for gene (codon optimized): **

<span style="background-color: yellow; font-family: Cambria,serif; font-size: 11pt;">Do Not Need this info for Spring <span style="font-family: Cambria,serif; font-size: 11pt;">-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
 * <span style="font-family: Cambria,serif; font-size: 11pt;">Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **
 * <span style="font-family: Cambria,serif; font-size: 11pt;">( ****<span style="color: green; font-family: Cambria,serif; font-size: 11pt;">link to DNA Works output text file - **<span style="font-family: Cambria,serif; font-size: 11pt;">that should be saved in your Google Docs folder after you did the primer design protocol)


 * <span style="font-family: Cambria,serif; font-size: 11pt;">Primer design results for 'tail' primers (this is just 2 sequences): **

<span style="background-color: white; font-family: Cambria,serif; font-size: 11pt;">[1] Nigou, J.; Dover, L. G.; Besra, G. S., Purification and Biochemical Characterization of Mycobacterium tuberculosis SuhB, an Inositol Monophosphatase Involved in Inositol Biosynthesis†. //Biochemistry// **2002,** //41// (13), 4392-4398. <span style="background-color: white; font-family: Cambria,serif; font-size: 11pt;">[2] Mayo Clinic. Tuberculosis. http://www.mayoclinic.com/health/tuberculosis/DS00372/ DSECTION=risk-factors (accessed May 1, 2013)