5-O-(1-carboxyvinyl)-3-phosphoshikimate+phosphate-lyase+(Campylobacter jejuni)

Campylobacteriosis is an infectious disease caused by ingesting food or drink contaminated with a bacteria called Campylobacter. Campylobacteriosis is estimated to be the second most common foodborn illness, affecting 1.3 million people each year (according to the CDC). Complications with campylobacteriosis can cause chronic illnesses. For example, people infected with Campylobacter have been known to develop severe arthritis and Guillain-Barré Syndrome (causes acute paralysis and potentially respiratory problems). Chorismate synthase is part of the shikimate pathway, an essential pathway in C. Jejuni. The shikimate pathway, links metabolism of carbohydrates to the biosynthesis of aromatic compounds. Chorismate synthase catalyzes the conversion of ESPS to chorismate in the last step of the shikimate pathway. https://www.patricbrc.org/portal/portal/patric/Feature?cType=feature&cId=PATRIC.1121102.3.ARGD01000025.CDS.9220.10320.fwd Is it a monomer or multimer as biological unit **? (make prediction at** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): Multimer according to the PSA query. http://www.brenda-enzymes.org/enzyme.php?ecno=4.2.3.5 BRENDA shows inhibition of chorismate synthase in other bacteria species.
 * Target (protein/gene name):** Chorismate Synthase
 * NCBI Gene # or RefSeq#:** PATRIC ID fig|1121102.3.peg.129
 * Protein ID (NP or XP #) or Wolbachia#:** No protein ID listed on PATRIC
 * Organism (including strain):** //Campylobacter ////jejuni //
 * Etiologic Risk Group (see link below):** Category B Priority
 * / Disease Information (sort of like the Intro to your Mini __Research Write__ up):**
 * Link to TDR Targets page (if present):** No TDR page
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)**
 * Essentiality of this protein:** Chorismate synthase has been found to be essential to //C.// //jejuni// as it is involved in the synthesis of aromatic amino acids.
 * Complex of proteins?:** Chorismate synthase is involved in the shikimate pathway, though catalyses the final step independently. It uses flavin mononucleotide (FMN) as a cofactor.
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):**

http://www.brenda-enzymes.org/enzyme.php?ecno=4.2.3.5 Reaction schematic could not be found on BRENDA; the following schematic is from Wikipedia
 * EC#:** 4.2.3.5
 * Link to BRENDA EC# page:**



A spectrophotometric assay of chorismate synthase had been developed. It involves the photoreduction of FMN (flavin mononucleotide) mediated by oxalate. Chorismate formation is monitored at 275 nm. The additions (minus the enzyme) are 50 µM EPSP (substrate), 10 µM FMN (cofactor), 1 mM sodium oxalate, 50 mM phosphate, or bis-Tris propane buffer. Photoreduction was done by illumination by white light (250-W halogen bulb). Sigma Aldrich has Shikimate-3-phosphate (supposedly used in the enzyme assay in the research paper) for $230.50 for 1 mg. Flavin mononucleotide is available from Sigma Aldrich at $25.00 for 100 mg. http://www.sigmaaldrich.com/catalog/product/sigma/e0377?lang=en®ion=US (substrate) -Provided by Sigma; E0377; $114 for 5 mg http://www.sigmaaldrich.com/catalog/product/aldrich/cds020791?lang=en®ion=US (cofactor) -Provided by Sigma; CDS020791; $25 for 100 mg http://www.sciencedirect.com/science/article/pii/S0003269784713091
 * 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**

-- PDB # or closest PDB entry if using homology model: 1SQ1
 * Structure (PDB or Homology model)**


 * Current Inhibitors:** No current inhibitors for chorismate synthesis in //C.// //jejuni//
 * Expression Information (has it been expressed in bacterial cells):** Has been expressed in //E. coli// according to BRENDA and research papers.
 * Purification Method :** Purified through centrifugation, fractation with ammonium sulfate, DEAE Sephacel chromatography, cellulose phosphate chromatography, and phenyl sepharose chromatography.


 * Image of protein (PyMol with features delineated and shown separately):**

MNTFGTRLKFTSFGESHGVAVGCIIDGMPAGVKFDEEFLQNELDKRKGGSKFATPRKESDKAQVLSGVFEGYTTGHPIAI VVFNENAHSKDYDNLKDLFRPAHADFTYFYKYGIRDHRGGGRSSARESVARVAGGAVAAMLLREFDICVQSGVFGVGTFV SNLKEEEFDFEFAKKSEIFCLDPKLESDFKNEILNARNSKDSVGAAVFTKVSGMLIGLGEVLYDKLDSKLAHALMGINAV KAVEIGEGINASKMRGSCNNDALKDGKFLSNHSGGILGGISNGENLILKTYFKPTPSIFAKQESIDKFGNNLKFELKGRH DPCVGVRGSVVASAMVRLVLADCLLLNASANLNNLKNAYGLKLEHHHHHH
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**
 * length of your protein in Amino Acids** 370
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website** 40316 kDa.
 * Molar Extinction coefficient of your protein at 280 nm wavelength:** 12295 (M-1)(cm-1)


 * TMpred graph Image** ( @http://www.ch.embnet.org/software/TMPRED_form.html ). Input your amino acid sequence to it.



ttgaatacttttggagtaaaacttagacttacaacttttggagaaagccatggcatagca attggtggcgtgcttgatggttttcctgctggagtaaagatagattttgattttttacaa aatgaacttgataaaagaaaacctagctcaaaatttgctaccaaaagaaaagaaagtgat aaagtagaagttttaagcggagtttttgaagggcttagcacgggaactccaattggtttt attataaaaaacgaagatcaaaaaagcaaagactatggaaatttaaaagatatttttaga ccaggacatgcagactatacatatttttataaatacggtataagagattatagaggtggt ggtagaagtagcgcaagagaaagtgcgataagagttgctggtggggcattttgccaaatg cttttaaatgagtttaaaataagtgttgaaagtggagtttttagtgttggggaagtaaat tttaacgctgattttaacgaggagtttaaaaaagggaatttagactttgaatttgctaaa acttcggaaattttctcactttttaaaaattttgatgaagagtttaaatctgaaatatta aaagccaaaaactctcataatagtgttggagcctcagttgtaactatcataaaaaactcc cccataggacttggagaggttttatatgataaatttgatgcaagattagtagctgctatg atggggataaatgctgtaaaagctgttgaaattggaaatggaataaaatcagccaaaatt tatggcgatgaaaacaatgatgaaatttcaaaaaatggttttttgacaaataattcaggt ggaattttgggtggcattacaaacggcgatgatattttaataaaatcatattttaaacca acaccatcaatttttttagaacaaaatactataaatttaagtggagaagatgaaatttgt aagctcaaaggaaggcatgatccttgcgttggaactagaggaagtgtagttgcaactgct atggcaaggctagttacagctgatatgttactactaaatacaagttcaaatttacaaaat cttaaaaaaatttatggctaa
 * CDS Gene Sequence (paste as text only):**
 * GC% Content for gene:** 31.97%
 * 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)

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