TargetSp17+-+Nitric-oxide+synthase (Bacillus+anthracis)


 * Target (protein/gene name):** nitric-oxide synthase, b. anthracis

***NCBI Gene # or RefSeq#:** NC_003997.3 Is it a monomer or multimer as biological unit ** ? (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): multimer in humans
 * *Protein ID (NP or XP #) or Wolbachia#: **
 * *Organism (including strain): ** bacillus anthracis
 * Etiologic Risk Group: ** Groups at risk include individuals who handle animals, like veterinarians, travelers and mail handlers. Vaccines can be provided to anyone who could possibly be exposed.
 * */ Disease Information: **Anthrax is a serious disease that is not contagious. This disease can affect animals along with humans. When spores from soil enter the body system, the bacteria begins to reproduce and the production of toxins begins. Anthrax is typically found in Central and South America, sub-Saharan Africa, central and southwestern Asia, southern and eastern Europe and the Caribbean. Anthrax can be cutaneous, inhalation, gastrointestinal and injection. If the spore entered through the skin, blisters could arise along with swelling. If inhaled, one can get chills, a cough and nausea. If the disease is gastrointestinal, one can feel a sore throat, nausea and vomiting, and stomach pain. Lastly, if the spores entered through injection, abscesses can occur along with blisters and swelling. Usually a diagnosis can occur after a chest x-ray, or some testing (blood, skin sample, etc.).
 * Link to Gene Database page: **https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2242674/
 * Essentiality of this protein: ** This protein is essential to bacteria. This is determined by the fact that pathogenic bacteria use their own nitric acid as a defense. ( https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2242674/)
 * Complex of proteins?: ** In humans, there are 4 amino acid chains and 15 ligands, making it complex.
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): n/a **


 * *EC#: **1.14.13.39
 * Link to BRENDA EC# page: **http://brenda-enzymes.org/enzyme.php?ecno=1.14.13.39&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Bacillus+anthracis&show_tm=0#SUBUNITS

Complicated assay - Dr. B 0606017

**---** Reagent: L arginine- 1 kg= $88.00
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * -- ** http://www.sigmaaldrich.com/catalog/search?interface=All&term=nitric+oxide+synthase&N=14572942+0+4294961277&mode=partialmax&focus=product&lang=en®ion=US
 * -- ** http://www.sigmaaldrich.com/catalog/substance/larginine174207479311?lang=en®ion=US

-- PDB #: 4NOS
 * Structure (PDB or Homology model) **

HUMAN INDUCIBLE NITRIC OXIDE SYNTHASE WITH INHIBITOR

10 20 30 40 50 MLSEKTIEIV KSTVPLLQEK GVEITTRFYE ILFSEHPELL NIFNHTNQKK 60 70 80 90 100 GRQQQALANA VYAAATYIDN LEAIIPVVKQ IGHKHRSLGI KAEHYPIVGT 110 120 130 140 150 CLLRAIKEVA GAPDEVLNAW GEAYGVIADA FISIEAEMYE EAAHKEGGWK 160 170 180 190 200 DFRNFVVVKK VKESDVITSL YLKPEDGGKV SSFIPGQYVT VQINIEGETY 210 220 230 240 250 THNRQYSLSD APGKEYYRIS VKKEKGVDTP DGKVSNYLHD HVKEGDMLPV 260 270 280 290 300 SAPAGDFVLN MDSTLPVVLI SGGVGITPMM SMLNTLIEQD SKRNVCFVHA 310 320 330 340 350 AINSNTHAMK EHVEAVDNEY EQVKAYTCYS APTEKDLEMK NFDKEGFVER 360 370 380 390 400 EWLQTIIPTT EAEFYFCGPV PFMKHINAVL TDLGVKQEHI HYEFFGPAAS
 * Current Inhibitors: **imidazole, NG-methyl arginine
 * Expression Information (has it been expressed in bacterial cells): ** Nitric oxide synthase has been in expressed and "functional in expression of the C-terminally His6-tagged soluble enzyme in Escherichia coli strain BL21"
 * Purification Method: ** Enzyme can be purified by nickel affinity chromatography
 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

**Molecular Weight** **of your protein:** 44765.94 **Molar** **Extinction coefficient** **of your protein at 280 nm wavelength:** 43570
 * *length of your protein in Amino Acids: 400 **
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.

atgctgagcgaaaaaaccattgaaattgtgaaaagcaccgtgccgctgctgcaggaaaaa ggcgtggaaattaccacccgcttttatgaaattctgtttagcgaacatccggaactgctg aacatttttaaccataccaaccagaaaaaaggccgccagcagcaggcgctggcgaacgcg gtgtatgcggcggcgacctatattgataacctggaagcgattattccggtggtgaaacag attggccataaacatcgcagcctgggcattaaagcggaacattatccgattgtgggcacc tgcctgctgcgcgcgattaaagaagtggcgggcgcgccggatgaagtgctgaacgcgtgg ggcgaagcgtatggcgtgattgcggatgcgtttattagcattgaagcggaaatgtatgaa gaagcggcgcataaagaaggcggctggaaagattttcgcaactttgtggtggtgaaaaaa gtgaaagaaagcgatgtgattaccagcctgtatctgaaaccggaagatggcggcaaagtg agcagctttattccgggccagtatgtgaccgtgcagattaacattgaaggcgaaacctat acccataaccgccagtatagcctgagcgatgcgccgggcaaagaatattatcgcattagc gtgaaaaaagaaaaaggcgtggataccccggatggcaaagtgagcaactatctgcatgat catgtgaaagaaggcgatatgctgccggtgagcgcgccggcgggcgattttgtgctgaac atggatagcaccctgccggtggtgctgattagcggcggcgtgggcattaccccgatgatg agcatgctgaacaccctgattgaacaggatagcaaacgcaacgtgtgctttgtgcatgcg gcgattaacagcaacacccatgcgatgaaagaacatgtggaagcggtggataacgaatat gaacaggtgaaagcgtatacctgctatagcgcgccgaccgaaaaagatctggaaatgaaa aactttgataaagaaggctttgtggaacgcgaatggctgcagaccattattccgaccacc gaagcggaattttatttttgcggcccggtgccgtttatgaaacatattaacgcggtgctg accgatctgggcgtgaaacaggaacatattcattatgaattttttggcccggcggcgagc
 * *CDS Gene Sequence (paste as text only): **

ATG CTT AGT GAA AAA ACG ATT GAA ATC GTG AAG TCA ACA GTG CCA CTG TTG CAA GAG AAG GGA GTA GAG ATT ACC ACG CGC TTT TAC GAG ATT TTA TTC TCC GAA CAC CCC GAG TTA TTG AAT ATT TTC AAT CAC ACG AAC CAG AAG AAG GGG CGT CAG CAA CAA GCG CTT GCC AAC GCA GTG TAC GCA GCA GCT ACC TAC ATC GAT AAC CTG GAG GCC ATT ATC CCT GTA GTT AAA CAA ATC GGG CAC AAG CAT CGC TCA CTG GGT ATC AAG GCA GAG CAC TAC CCC ATC GTG GGG ACT TGT TTA TTA CGT GCA ATT AAA GAG GTA GCG GGT GCA CCC GAT GAG GTG TTA AAT GCA TGG GGG GAA GCA TAT GGT GTC ATT GCA GAC GCG TTC ATC TCA ATC GAG GCC GAA ATG TAT GAG GAA GCC GCG CAT AAG GAA GGG GGT TGG AAA GAC TTC CGC AAC TTC GTT GTA GTG AAA AAA GTG AAG GAA TCC GAT GTT ATT ACC TCA TTA TAC TTA AAG CCC GAA GAT GGT GGA AAA GTT TCT AGT TTC ATT CCT GGT CAG TAT GTA ACT GTC CAG ATC AAT ATC GAA GGC GAA ACT TAT ACA CAC AAT CGT CAA TAT TCT TTG AGC GAC GCT CCG GGG AAG GAA TAT TAC CGC ATC AGC GTG AAA AAA GAG AAG GGA GTC GAC ACA CCC GAT GGT AAA GTT TCA AAC TAC CTG CAC GAT CAC GTA AAG GAA GGG GAC ATG CTT CCA GTT AGT GCT CCA GCA GGC GAC TTT GTG CTT AAC ATG GAC TCT ACG TTA CCA GTG GTT TTA ATT TCT GGA GGA GTT GGA ATT ACC CCC ATG ATG AGC ATG CTG AAT ACC TTA ATC GAA CAG GAC TCT AAA CGC AAC GTC TGT TTT GTC CAT GCG GCC ATC AAC AGT AAT ACA CAT GCG ATG AAG GAG CAT GTT GAG GCG GTC GAC AAC GAA TAC GAG CAA GTT AAG GCC TAT ACG TGC TAC TCC GCG CCT ACA GAG AAG GAT TTG GAG ATG AAA AAC TTC GAT AAA GAG GGT TTT GTC GAA CGT GAG TGG TTA CAG ACG ATC ATC CCT ACC ACG GAA GCT GAA TTT TAC TTC TGC GGT CCG GTG CCG TTT ATG AAG CAT ATT AAC GCG GTT CTT ACC GAC CTT GGG GTG AAA CAA GAG CAT ATC CAT TAT GAA TTT TTC GGA CCT GCC GCA AGC
 * *GC% Content for gene: 50.08% **
 * *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): **


 * *GC% Content for gene (codon optimized): ** 46.4 %

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)

**  also known as RB-1605-P1ABX ||   ||
 * Primer design results for 'tail' primers (this is just 2 sequences): **
 * RB-1605-P1ABX