Target+-+3-oxoacyl-+(acyl-carrier-protein)+reductase+(Mycobacterium+tuberculosis)

Tuberculosis is a potentially fatal disease that is caused by the bacteria //Mycobacterium tuberculosis.// In the past, tuberculosis was the leading cause of death in the United States. Although this is no longer the case due to vaccinations and immunizations, the threat of tuberculosis is still a reality. African-Americans, pregnant women, and internal travelers are all at higher risks to contracting tuberculosis based on statistical data. Complications with eradicating tuberculosis include the development of drug-resistance from poor quality drugs or people not completing treatment. Tuberculosis can also stay inactive in the body and become activated causing the normal symptoms of tuberculosis to arise. This has been termed latent tuberculosis infection. [|[1]] -Gene/Ortholog: mtu1373 (OG4_10098); Phenotype: essential; Source study: nmpdr
 * *Target (protein/gene name): ** 3-oxoacyl-[acyl-carrier-protein] reductase / fabG1
 * *NCBI Gene # or RefSeq#: ** 886551
 * *Protein ID (NP or XP #) or Wolbachia#: **NP_215999.1
 * *Organism (including strain): **//Mycobacterium tuberculosis H37Rv//
 * Etiologic Risk Group (see link below): ** Risk Group 3
 * *Background/Disease Information (sort of like the Intro to your Mini Research Write up): **
 * Essentiality of this protein: **

-Gene/Ortholog: eco1057 (OG4_10098); Phenotype: essential; Source study: gerdes


 * Complex of proteins?: ** No
 * Druggable Target: ** Yes, 0.7 druggability index


 * *EC#: ** 1.1.1.100
 * Link to BRENDA EC# page: ** [] [2]
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic

[|Potassium phosphate], [|NADPH], [|acetoacetyle-N-acetylcysteamine] [3] For same protein in different organism (Persea americana) [|Sheldon, P.S.; Kekwick, R.G.O.; Sidebottom, C.; Smith, C.G.; Slabas, A.R., 3-Oxoacyl-(acyl-carrier protein) redictase from avocado (Persia americana) fruit mesocarp. Biochem. J. 1990, 271, 713-720.] [4] Potassium phosphate - $20.80 NADPH - $99.90 acetoacetyle-N-acetylcysteamine - $70.10
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): ** Spectrophotometric; Reagents: 100 mM Potassium phosphate (pH 7.0), NADPH (100 uM), acetoacetyle-N-acetylcysteamine (5mM)
 * -- link to Sigma (or other company) page for assay or assay reagents (substrates) **
 * -- link (or citation) to paper that contains assay information **
 * -- List cost and quantity of substrate reagents and supplier **

-- PDB # or closest PDB entry if using homology model: 1UZL 1UZM 1UZN 2NTN
 * Structure Available (PDB or Homology model) **

MTATATEGAKPPFVSRSVLVTGGNRGIGLAIAQRLAADGHKVAVTHRGSGAPKGLFGVEVDVTDSDAVDRAFTAVEEHQG PVEVLVSNAGLSADAFLMRMTEEKFEKVINANLTGAFRVAQRASRSMQRNKFGRMIFIGSVSGLWGIGNQANYAASKAGV IGMARSIARELSKANVTANVVAPGYIDTDMTRALDERIQQGALQFIPAKRVGTPAEVAGVVSFLASEDASYISGAVIPVD GGMGMGH
 * Current Inhibitors: ** 14 in the species //Plasmodium falciparum// but none in //Mycobacterium tuberculosis// on The Binding Database
 * Expression Information (has it been expressed in bacterial cells): **[|Yes, in E. coli] [6]
 * Purification Method: ** Column Chromatography ** - ** " Induced recombinant bacteria were washed with cold 50 mM potassium phosphate buffer pH 7±8 (buffer A), and resuspended in 4 ml of the same buffer supplemented with 500 mM NaCl and 5 mMimidazole. Cells were broken by one freezing}thawing cycle at®70 °C, in the presence of protease inhibitors [aprotinin, soybean trypsininhibitor, TLCK (N-p-tosyl-lysine chloromethyl ketone), pepstatin, leupeptin and PMSF] and 0±5 mg lysozyme ml−". Nucleic acids were removed by DNase I (5 lg ml−") and RNase A (10 lg ml−") treatments in the presence of MgCl# (10 mM) at 4 °C for 15 min. After centrifugation at 44000 g for 15 min, the supernatant supplemented with 10% (v}v) glycerol was applied to a Ni-NTA Agarose (Qiagen) column (0±5 ml bed volume). After extensive washes by 5 mM then 50 mM imidazole in buffer A supplemented with 500 mM NaCl, MabA was eluted with 175 mM imidazole in buffer A supplemented with 500 mM NaCl. Fractions containing the His-tagged MabA (H-MabA) were identi®ed by SDS-PAGE, pooled, dialysed twice against 50 vols 50% (v}v) glycerol in 50 mM potassium phosphate buffer, pH 7±2, and stored at ®20 °C." (Same paper as expression linked above). [6]
 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

[8] GTGACTGCCACAGCCACTGAAGGGGCCAAACCCCCATTCGTATCCCGTTCAGTCCTGGTTACCGGAGGAAA CCGGGGGATCGGGCTGGCGATCGCACAGCGGCTGGCTGCCGACGGCCACAAGGTGGCCGTCACCCACCG TGGATCCGGAGCGCCAAAGGGGCTGTTTGGCGTCGAATGTGACGTCACCGACAGCGACGCCGTCGATCGC GCCTTCACGGCGGTAGAAGAGCACCAGGGTCCGGTCGAGGTGCTGGTGTCCAACGCCGGCCTATCCGCGG ACGCATTCCTCATGCGGATGACCGAGGAAAAGTTCGAGAAGGTCATCAACGCCAACCTCACCGGGGCGTTC CGGGTGGCTCAACGGGCATCGCGCAGCATGCAGCGCAACAAATTCGGTCGAATGATATTCATAGGTTCGGT CTCCGGCAGCTGGGGCATCGGCAACCAGGCCAACTACGCAGCCTCCAAGGCCGGAGTGATTGGCATGGCC CGCTCGATCGCCCGCGAGCTGTCGAAGGCAAACGTGACCGCGAATGTGGTGGCCCCGGGCTACATCGACA CCGATATGACCCGCGCGCTGGATGAGCGGATTCAGCAGGGGGCGCTGCAATTTATCCCAGCGAAGCGGGT CGGCACCCCCGCCGAGGTCGCCGGGGTGGTCAGCTTCCTGGCTTCCGAGGATGCGAGCTATATCTCCGGTG CGGTCATCCCGGTCGACGGCGGCATGGGTATGGGCCACTGA [9]  1 ATGACCGCGACGGCGACCGAAGGTGCGAAACCGCCGTTCGTTTCCCGTAGCGTTCTCGTT  61 ACGGGTGGTAATCGTGGCATTGGTCTGGCTATCGCCCAACGTCTCGCTGCCGACGGTCAC 121 AAGGTTGCGGTTACTCACCGTGGTTCTGGTGCGCCGAAAGGTCTGTTCGGTGTTGAGTGC 181 GACGTTACTGATTCTGACGCGGTGGATCGCGCTTTCACCGCTGTTGAAGAACACCAGGGT 241 CCGGTAGAAGTACTCGTATCTAACGCCGGTCTGTCTGCGGACGCTTTCCTGATGCGTATG 301 ACTGAAGAAAAATTCGAAAAAGTTATTAACGCGAACCTGACCGGTGCGTTCCGTGTTGCG 361 CAGCGTGCGTCTCGTTCTATGCAGCGTAACAAGTTTGGTCGTATGATTTTCATCGGTTCT 421 GTTTCTGGTTCTTGGGGCATCGGCAACCAGGCGAATTATGCGGCGAGCAAAGCCGGTGTG <span style="background-color: #ffffff; font-family: 'Courier New',Courier,monospace; font-size: 14px;">481 ATCGGTATGGCGCGTTCCATCGCGCGTGAACTGTCTAAAGCTAATGTTACTGCTAACGTT <span style="background-color: #ffffff; font-family: 'Courier New',Courier,monospace; font-size: 14px;">541 GTCGCGCCTGGCTACATCGACACGGATATGACTCGCGCGCTCGACGAGCGTATCCAGCAG <span style="background-color: #ffffff; font-family: 'Courier New',Courier,monospace; font-size: 14px;">601 GGTGCGCTGCAATTCATCCCGGCCAAACGTGTTGGTACCCCGGCAGAAGTTGCGGGTGTT <span style="background-color: #ffffff; font-family: 'Courier New',Courier,monospace; font-size: 14px;">661 GTCTCTTTTCTGGCGTCTGAAGACGCATCTTACATTTCCGGTGCCGTTATCCCGGTCGAC <span style="background-color: #ffffff; font-family: 'Courier New',Courier,monospace; font-size: 14px;">721 GGTGGCATGGGTATGGGTCACTAA
 * *length of your protein in Amino Acids: ** 247 residues
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: **25.7192 kDa [7]
 * Molar Extinction coefficient of your protein at 280 nm wavelength: **9970
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
 * *CDS Gene Sequence (paste as text only): **
 * *GC% Content for gene: ** [|64.4%] [10]
 * *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): **


 * *GC% Content for gene (codon optimized): 55.4% **

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)


 * Primer design results for 'tail' primers (this is just 2 sequences): **

1. [] 2. [] 3. [] 4. [] 5. [] 6. [] 7. [] 8. [] 9. [] 10. [] 11. []
 * References: **