TargetSp17+-+FabI+enoyl-acyl+carrier+protein+reductase+(Francisella+tularensis)

// Francisella tularensis // is responsible for the rare disease, Tularemia. It is most commonly found in rodents such as rabbits and hares, but it can be transmitted in humans. The disease can be spread to humans via insect bites and direct exposure to an infected animal. The highly contagious disease mainly attacks the skin, eyes, lymph nodes, and lungs. If left untreated, the disease can be fatal. There are many different types of the disease including Ulceroglandular tularemia, Oculoglandular tularemia, Oropharyngeal tularemia, and Pneumonic tularemia. The most common variant of this disease is the Ulceroglandular type. Signs and symptoms of this type include: a skin ulcer that forms at the site of infection, swollen and painful lymph glands, fever, chills, headache, and exhaustion. The common symptom among all the types of the tularemia includes ulcers at the sight of the infection. The dangerous thing about this disease is the fact that it can be used for bioterrorism. The fact that it’s highly contagious and infective means that it can pose a large threat if used as a weapon. [] [] - The Francisella tularensis FabI enoyl-acyl carrier protein reductase (FtFabI) gene is essential to bacterial viability and is expressed during infection. FtFabI is critical for fatty acid biosynthesis within the Francisella tularensis organism. The inhibition of this protein results in the loss of viability of the organism even if supplements are used. It has also ecome known that the protein is highly active during the process of infection. Without the gene, the organism will not be able to properly function normally or in times of infection. Is it a monomer or multimer as biological unit **? (make prediction at** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): Monomer []
 * Target (protein/gene name):** FabI enoyl-acyl carrier protein reductase (//Francisella tularensis)//
 * NCBI Gene # or RefSeq#:** 3191864/NC_006570.2
 * Protein ID (NP or XP #) or Wolbachia#:**
 * Organism (including strain):** //Francisella tularensis//
 * Etiologic Risk Group (see link below):** RG2
 * / Disease Information (sort of like the Intro to your Mini __Research Write__ up):**
 * Link to TDR Targets page (if present): N/A**
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)**
 * Essentiality of this protein:**
 * Complex of proteins?:**
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):**

[] http://www.med.upenn.edu/ostaplab/documents/NADHLinkedAssayforATPaseMeasurements.pdf http://www.jbc.org/content/256/13/6583.full.pdf [] [] 1 mg NADH (Sigma, N0786) - $139.50 -- PDB # or closest PDB entry if using homology model: 2JJY -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB Query Coverage: Max % Identities: % Positives Chain used for homology:
 * EC#:** EC 1.3.1.9
 * Link to BRENDA EC# page:**
 * --** Show screenshot of BRENDA enzyme mechanism schematic
 * Figure 1.** BRENDA enzyme mechanism schematic of the enoyl-[acyl-carrier-protein] reductase (NADH)
 * 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 #**
 * Structure (PDB or Homology model)**

[] The protein is made up of 4 identical chains. Below is the sequence for a single chain: MGSSHHHHHHSSGLVPRGSHMGFLAGKKILITGLLSNKSIAYGIAKAMHREGAELAFT YVGQFKDREKLCAEFNPAAVLPCDVISDQEIKDLFVELGKVWDGLDAIVHSIAFAPRDQ LEGNFIDCVTREGFSIAHDISAYSFAALAKEGRSMMKNRNASMVALTYIGAEKAMPSYN TMGVAKASLEATVRYTALALGEDGIKVNAVSAGPIKTLAASGISNFKKMLDYNAMVSPL KKNVDIMEVGNTVAFLCSDMATGITGEVVHVDAGYHCVSMGNVL 280 Amino Acids x 4 = 1,120 Amino Acids 29968.54 Da x 4 = 119,874.16 Da ATGGGTTTTCTAGCAGGAAAAAAAATATTAATCACTGGACTTTTAAGTAATAAGTCAATTGCATATGGTA TTGCTAAAGCTATGCATAGAGAGGGAGCCGAGCTTGCTTTTACTTATGTTGGACAGTTCAAAGATAGA GTGGAAAAATTATGTGCAGAATTTAATCCAGCTGCAGTTTTGCCTTGCGATGTGATTTCTGATCAAGA GATTAAGGATTTATTTGTAGAGCTAGGTAAAGTTTGGGATGGTCTAGATGCCATAGTTCATTCTATAGC TTTTGCACCGCGTGATCAGTTAGAAGGTAACTTTATTGACTGTGTAACTCGCGAGGGTTTTAGTATCG CTCATGATATTAGTGCCTATTCTTTTGCAGCATTAGCTAAAGAAGGTCGTAGTATGATGAAAAATCGTAA TGCTTCTATGGTAGCACTTACTTATATTGGAGCAGAAAAAGCTATGCCAAGTTACAATACTATGGGAGT TGCTAAAGCATCTCTAGAAGCTACAGTTAGATATACAGCGTTAGCTTTAGGTGAGGATGGTATCAAGG TAAATGCTGTATCAGCTGGTCCTATCAAAACTCTGGCAGCTTCTGGTATATCAAACTTCAAGAAGATGC TTGATTATAATGCTATGGTTTCTCCACTTAAGAAAAATGTTGATATTATGGAAGTTGGTAATACTGTAGCG TTTTTATGTTCAGATATGGCAACTGGTATCACTGGAGAAGTTGTCCATGTTGATGCTGGATATCATTGTG TGTCTATGGTAATGTTCTTTAA
 * Current Inhibitors: Triclosan**
 * Expression Information (has it been expressed in bacterial cells):** Has been expressed in E. coli
 * Purification Method :** Nickel Column Chromatography
 * Image of protein (PyMol with features delineated and shown separately):**
 * Figure 2.** PyMOL representation of the fabI enoyl-ACP reductase protein found in Francisella tularensis. The chains are colored separately and labeled (Green – A, Blue – B, Pink – C, Yellow – D). The protein is seen in its cartoon form. From 2JJY PDB file.
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**
 * 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:** 17670 mol-1 cm-1
 * TMpred graph Image** ( @http://www.ch.embnet.org/software/TMPRED_form.html ). Input your amino acid sequence to it.
 * Figure 3.** TMpred output graph for fabI enoyl-acyl carrier protein reductase found in Francisella tularensis
 * CDS Gene Sequence (paste as text only):**
 * GC% Content for gene:** 37.037%
 * CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):**
 * GC% Content for gene (codon optimized):**

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)