Fab+G+3-ketoacyl-(acyl-carrier-protein)+reductase


 * Target (protein/gene) name: Fab G 3-ketoacyl-(acyl-carrier-protein) reductase **


 * NCBI Gene # or RefSeq#: 883564 **


 * Protein ID (NP or XP#) or Wolbachia#: NP_221114.1 **


 * Organism (including strain): Rickettsia prowazekii **


 * Etiologic Risk Group: Appendix B-III-A. Risk Group 3 (RG3) - Bacterial Agents Including Rickettsia **


 * Background/Disease information:**
 * The disease involved with this gene/protein (Fab G-ketoacyl-(acyl-carrier-protein) reductase) is epidemic typhus, which is caused by Rickettsia prowazekki bacteria. This disease spreads to humans through lice or ticks and can be found on the flying squirrel. Although uncommon in the US, several cases of typhus have been reported in Texas, in particular in Austin. Since the US is hygienically advanced, it is less prone to exposure and spread of typhus; but areas of the world with lower hygiene and cold temperature, like certain third world countries, are much more likely to have a large epidemic due to typhus. After contraction, the disease may be dormant for years before it re-activates. Death may occur, if left untreated, in ten to sixty percent of patients. Symptoms include the following: **
 * ** severe headache **
 * ** high fever **
 * ** rash that begins on the back or chest/spreads **
 * ** confusion **
 * ** stupor and seeming out of touch with reality **
 * ** low blood pressure **
 * ** eye sensitivity to bright lights **
 * ** severe muscle pain **
 * Common treatments include antibiotics like tetracycline and the disease can be prevented by bathing, avoiding infested areas, and sterilizing infected clothing. **


 * Link to TDR Targets page: not applicable**

[]
 * Link to Gene Database page**


 * Essentiality of this protein**
 * super essential in biosynthesis (fatty acids) **


 * structure of 3-ketoacyl-(acyl-carrier-protein) reductase from Rickettsia prowazekii at 2.25 A res **
 * []


 * Complex of proteins: tetramer **


 * Druggable Target (list, number, or cite evidence from a paper/database showing druggable in another organism):**
 * ** Inhibition of Plasmodium falciparum Fatty Acid Biosynthesis: Evaluation of FabG, FabZ, and FabI as Drug Targets for Flavonoids **
 * ** Structure of 3-ketoacyl-(acyl-carrier-protein) reductase from Rickettsia prowazekii at 2.25A° resolution **
 * []


 * EC#: 1.1.1.100 **


 * Link to BRENDA EC#:**


 * Enzyme Assay Information:**
 * (3R)-3-hrdroxyacyl-[acyl-carrier protein] plus NADP+; spectrophotometric **


 * Substrate Info-100113**


 * NADPH**
 * //Preparation Instructions //**
 * β-NADPH is soluble in 0.01 M sodium hydroxide (50 mg/ml), yielding a clear, light yellow solution **.


 * //Storage/Stability //**
 * It is recommended to store Products N1630, N7505, and N6505 desiccated at –20 °C protected from light. Product N9910 can be stored at room temperature. The normal impurities and/or decomposition products are β-NADP and monophosphoadenosine 5′- diphosphoribose. It is recommended to prepare solutions fresh and use promptly, unless you are sure this is an unnecessary precaution for your work. However, it has been reported that a 0.5 mM solution in 0.02 M NaOH (pH 12.3) showed no loss of purity in a week at 4 °C or −85 °C, but a 13% loss at –20 °C.3 **


 * Structure Available (PDB or Homology model)**
 * PDB: 3F91 **


 * Current Inhibitors**
 * Flavanoids, EGCG in green tea **


 * Expression Information**
 * in E. Coli cells **


 * Purification Method:**
 * Nickel + FPLC **


 * Image of protein**


 * Amino Acid Sequence**
 * MIDLTGKTSLITGASSGIGSAIARLLHKLGSKVIISGSNEEKLKSLGNALKDNYTIEVCNLANKEECSNLISKTSNLDILVCNAGITSDTLAIRMKDQDFDKVIDINLKANFILNREAIKKMIQKRYGRIINISSIVGIAGNPGQANYCASKAGLIGMTKSLSYEVATRGITVNAVAPGFIKSDMTDKLNEKQREAIVQKIPLGTYGIPEDVAYAVAFLASNNASYITGQTLHVNGGMLMVX **


 * Length of protein:**
 * 242 **


 * Molecular Weight in kiloDaltons:**
 * 25870.1 kD **


 * Molar Extinction coefficient of your protein at 280 nm wavelength:**
 * 10680 M-1 cm-1, 10430 M-1 cm-1 **


 * CDS Gene Sequence**
 * CAAACAATATTTAAAAGAACATCCACAAATTTCCAACGAAATTGAGAAAATAATACGAGAAAAATCGTCAGCAATTACTAATATAAATCTTGATCAAACGGAGGAATAAAATGATTGATTTAACAGGTAAAACTTCCTTAATTACAGGTGCTTCAAGTGGCATAGGAAGTGCCATAGCGCGGTTACTACATAAACTCGGGAGCAAGGTAATTATTAGCGGTAGTAATGAAGAAAAATTAAAATCACTTGGCAATGCTTTAAAAGATAATTATACCATAGAAGTATGTAATCTTGCGAATAAGGAAGAATGTAGTAATTTAATATCTAAAACATCAAATTTAGATATTTTAGTATGTAATGCCGGTATTACTAGTGATACACTCGCAATTAGAATGAAAGATCAAGATTTCGACAAAGTTATTGATATTAATTTGAAAGCCAATTTTATTTTAAATCGTGAAGCAATAAAAAAAATGATACAAAAAAGGTATGGACGCATTATTAACATATCTTCAATAGTAGGAATTGCAGGTAACCCAGGACAGGCTAATTATTGCGCTTCTAAAGCAGGTTTAATAGGTATGACCAAATCGCTATCTTACGAAGTTGCAACTAGAGGGATTACGGTTAATGCAGTAGCACCAGGATTTATTAAATCAGATATGACTGATAAACTCAATGAAAAACAACGAGAAGCAATAGTACAAAAAATTCCACTAGGTACTTATGGTATACCAGAAGACGTTGCATACGCAGTCGCATTTTTAGCAAGCAATAATGCATCATATATTACTGGTCAAACTCTACATGTAAACGGTGGTATGTTAATGGTATAAAATTTTATGCTAGCAAAAAATAATTTTTTGTGCTAGAAGTTCTTTTAAAGAGAAAATGCTTATGAATATTTATTTCAAATATTCATAGCAATGAAAATAAAATCTAAT **

**30.6 %**
 * GC% Content for gene:**


 * CDS Gene Sequence (codon optimized)**
 * ATGATTGACCTCACGGGCAAAACTTCTCTGATCACGGGTGCCAGCTCTGGTATCGGTTCTGCAATCGCGCGTCTGCTGCACAAACTGGGCTCTAAAGTTATTATCTCTGGTTCTAACGAAGAAAAACTGAAGTCTCTCGGTAACGCGCTCAAAGACAACTACACCATCGAAGTTTGTAACCTCGCGAACAAAGAAGAATGCAGCAATCTGATCTCTAAAACCTCTAATCTGGACATCCTGGTTTGCAATGCGGGCATCACCTCTGACACCCTGGCGATCCGTATGAAAGACCAGGACTTTGACAAAGTCATCGATATCAACCTGAAAGCGAACTTCATCCTGAACCGTGAAGCGATCAAGAAGATGATCCAGAAACGTTACGGTCGTATTATCAACATCTCCTCTATCGTTGGTATTGCGGGTAACCCAGGTCAGGCGAACTACTGCGCATCTAAAGCGGGCCTGATCGGCATGACCAAATCTCTGTCTTACGAAGTTGCGACCCGCGGTATCACCGTCAACGCCGTTGCGCCAGGTTTCATCAAATCTGACATGACTGACAAGCTGAATGAAAAACAGCGTGAGGCAATCGTTCAGAAAATCCCGCTGGGTACCTACGGTATTCCAGAAGACGTGGCTTACGCGGTTGCGTTCCTGGCGTCTAACAACGCGTCTTACATTACCGGTCAAACCCTGCATGTTAACGGTGGTATGCTGATGGTTTAA **


 * GC% Contect for gene:**
 * 48.6 % **


 * Primer Design results for pNIC-Bsa4 cloning:**
 * 1 ATGATTGACCTCACGGGCAAA 21 **
 * 2 GCAGAACCGATACCAGAGCTGGCACCCGTGATCAGAGAAGTTTTGCCCGTGAGGTCAATC 60 **
 * 3 GCTCTGGTATCGGTTCTGCAATCGCGCGTCTGCTGCACAAACTGGGCTCTAAAGTTATTA 60 **
 * 4 GAGAGACTTCAGTTTTTCTTCGTTAGAACCAGAGATAATAACTTTAGAGCCCAGTTTGTG 60 **
 * 5 TAACGAAGAAAAACTGAAGTCTCTCGGTAACGCGCTCAAAGACAACTACACCATCGAAGT 60 **
 * 6 ATCAGATTGCTGCATTCTTCTTTGTTCGCGAGGTTACAAACTTCGATGGTGTAGTTGTCT 60 **
 * 7 AAAGAAGAATGCAGCAATCTGATCTCTAAAACCTCTAATCTGGACATCCTGGTTTGCAAT 60 **
 * 8 CTTTCATACGGATCGCCAGGGTGTCAGAGGTGATGCCCGCATTGCAAACCAGGATGTCCA 60 **
 * 9 CCTGGCGATCCGTATGAAAGACCAGGACTTTGACAAAGTCATCGATATCAACCTGAAAGC 60 **
 * 10 CATCTTCTTGATCGCTTCACGGTTCAGGATGAAGTTCGCTTTCAGGTTGATATCGATGAC 60 **
 * 11 CGTGAAGCGATCAAGAAGATGATCCAGAAACGTTACGGTCGTATTATCAACATCTCCTCT 60 **
 * 12 GTTCGCCTGACCTGGGTTACCCGCAATACCAACGATAGAGGAGATGTTGATAATACGACC 60 **
 * 13 ACCCAGGTCAGGCGAACTACTGCGCATCTAAAGCGGGCCTGATCGGCATGACCAAATCTC 60 **
 * 14 CGTTGACGGTGATACCGCGGGTCGCAACTTCGTAAGACAGAGATTTGGTCATGCCGATCA 60 **
 * 15 GCGGTATCACCGTCAACGCCGTTGCGCCAGGTTTCATCAAATCTGACATGACTGACAAGC 60 **
 * 16 GATTTTCTGAACGATTGCCTCACGCTGTTTTTCATTCAGCTTGTCAGTCATGTCAGATTT 60 **
 * 17 TGAGGCAATCGTTCAGAAAATCCCGCTGGGTACCTACGGTATTCCAGAAGACGTGGCTTA 60 **
 * 18 TGTAAGACGCGTTGTTAGACGCCAGGAACGCAACCGCGTAAGCCACGTCTTCTGGAATAC 60 **
 * 19 CGTCTAACAACGCGTCTTACATTACCGGTCAAACCCTGCATGTTAACGGTGGTATGCTGA 60 **
 * 20 TTAAACCATCAGCATACCACCGTTAACAT 29 **


 * Primer design results for tail primers**


 * forward**
 * 5’ TACTTCCAATCCATGATTGACCTCACGGGCAA 3' **
 * reverse**
 * 5’ GTGGTATGCTGATGGTTTAACAGTAAAGGTGGATA 3’ **