Clostripain+(Clostridium+botulinum)


 * Target (protein/gene name): Clostripain**


 * NCBI Gene # or RefSeq#: 5397352**


 * Protein ID (NP or XP #) or Wolbachia#: 4YEC**


 * Organism (including strain): //Clostridium//** //**botulinum**//


 * Etiologic Risk Group (see link below): //2//**


 * / Disease Information (sort of like the Intro to your Mini __Research Write__ up):**
 * //Clostridium botulinum// is the bacteria that causes botulism. Botulism is an intestinal disease caused by undercooked food, entry into wounds, or inhalation of spores. Botulism is highly preventable, as food-bourns botulism dies at** **temperatures above 185 degrees F and most wound botulism is the product of street drug injections. Botulism is generally treated with antibiotics, which have a moderate success rate, leaving about a 5-10% mortality rate. Most issues with botulism arise from misdiagnosing the symptoms as a common virus, allowing the bacteria to grow strong enough that it isn't eradicated by antibiotics, or simple lack of antibiotic availability (non-issue in first-world countries).**


 * Link to TDR Targets page (if present):**

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 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.):**


 * Essentiality of this protein: pathogenesis**

Is it a monomer or multimer as biological unit **? (make prediction at** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): multimer (di-mer)


 * Complex of proteins?: no**

The best known of these is leupeptin (N- acetyl-L-leucyl-L-leucyl-D,L-argininaldehyde), which was originally isolated from Streptomyces exfoliates and inhibits a wide range of serine, cysteine and threonine-type peptidases, including trypsin, PACE4 calpain, clostripain and the trypsin-like activity of the proteasome (Aoyagi et al., 1969; Kembhavi et al., 1991; Kurinov and Harrison, 1996; Mains et al., 1997; Moldoveanu et al., 2004; Savory et al., 1993).
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):**


 * EC#: 3.4.22.8**

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 * Link to BRENDA EC# page:**


 * --** Show screenshot of BRENDA enzyme mechanism schematic
 * Enzyme schematic was not offered on BRENDA page

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 * 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**

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 * -- links to assay reagents (substrates) pages.**


 * --- List cost and quantity of substrate reagents, supplier, and catalog #**
 * Clostripain (Endoproteinase-Arg-C); **10 ug; LS02135; $83.00.

-- PDB # or closest PDB entry if using homology model: 4YEC
 * Structure (PDB or Homology model)**


 * Current Inhibitors:**
 * EDTA
 * Oxidizing agents
 * Sulfhydryl reagents (such as TLCK) (Porter //et al.// 1971)
 * Co2+, Cu2+, Cd2+, and heavy metal ions
 * Citrate, borate and Tris anions partially inhibit

Clostripain is expressed by the anaerobic bacteria //Clostridium histoliticum. //Clostripain has also been expressed and isolated from //Escherichia coli //and //Bacillus subtilis //.
 * Expression Information (has it been expressed in bacterial cells):**

cation-exchange chromatography and ultrafiltration
 * Purification Method :**

METPEPR TTRTILVYMM A NNSL <span style="color: #ff0000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">NSFASKNIESMIEG AT <span style="color: #ff0000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">SKNL <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">NG G <span style="color: #0000ff; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">NLIVY EEE EEEEEEE SS TTHHHHHHHH HHHHHH HH HHTT EEEEE <span style="color: #0000ff; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">YA P <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">AGS PP <span style="color: #0000ff; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">ELLRIKEE <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">NG <span style="color: #0000ff; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">VVKKIHLKD YE <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">K QN <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">SAD <span style="color: #ff0000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">PDVMRSVIGEVVSQ <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">Y EE SSS EE EEEEEETTEE EEEEEEE S TTSHHHHH HHHHHHHHHS P <span style="color: #0000ff; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">ADSYGLVLW <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">S H <span style="color: #0000ff; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">G T <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">AW LP <span style="color: #008000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">SD <span style="color: #ff0000; font-family: Arial; font-size: 14.666666666666666px; vertical-align: baseline;">YQNK LK EEEEEEEEE S B TT TT GGGG
 * Image of protein (PyMol with features delineated and shown separately):**
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**


 * length of your protein in Amino Acids**
 * 126**

<span style="background-color: #ffffff; font-family: Verdana,Geneva,Arial,sans-serif;">53.0 kDa
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website**

<span style="background-color: #ffffff; font-family: Verdana,Geneva,Arial,sans-serif;">87,890 cm -1<span style="background-color: #ffffff; font-family: Verdana,Geneva,Arial,sans-serif;"> M -1
 * Molar Extinction coefficient of your protein at 280 nm wavelength:**


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

<span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">GCTATAATCAATGCCATATATATAAAGATTAAAGTAATCTTTTGTATCATCTAATAATGG <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">AGTATCAGCTTTATTAATTAATTGCTTTTTTAAAAAATTATTTGTAAGTTAGTATACTCT <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">TCTACTAAATAGTTAAATTCAGCTTTTCATTTTTCCATCACTACCATAATTACGTAATCA <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">TTATATCATAGAATGCTTTTACCACGTAATATAAAAAGCAAGAGTCCTATATTAAATCCT <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">TGAACCTAAAGACCTAATGTAGATTTTTAACAAAACAAATGATTGATCACTTAATATATC <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">TAACAGTTCAGATAACTTCCTTTTTGAGATTGTAATATATAAGTAAATTCAAAGACCATA <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">AAATATACCCCCCGATTGTAATGCTTGAATATTAATTATAATGAAATTATAGTTTAAATT <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">AAAAAAATAATTTTTTTATGAAGTTTAAGAAGGATTATCCTAAACATTATAGAATATATT <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">AAATATAAAAATAATGTAAAACTTTAAATAATAACTCTTATAATGGTACATAATGTATTT <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">ATATGGATTAAATTTTAATTTATAGATGCTATAAAACTATAAATTAAATTTAAAATATAC <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">TTTTGGAGGTATGTATTATGTTAAGAAGAAAAGTATCAACACTATTAATGACAGCTTTGA <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">TAACTACTTCATTTTTAAATTCCAAACCCGTATATGCAAATCCAGTAACTAAATCCAAGG <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">ATAATAACTTAAAAGAAGTACAACAAGTTACAAGCAAGAGTAATAAAAACAAAAATCAAA <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">AAGTAACTATTATGTACTATTGCGACGCAGACAATAACTTGGAAGGAAGTCTATTAAATG <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">ATATCGAGGAAATGAAAACAGGATATAAGGATAGTCCTAATTTAAATTTAATTGCTCTTG <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">TAGACAGATCCCCAAGATATAGCAGTGACGAAAAAGTTTTAGGTGAAGATTTTAGTGATA <span style="color: #222222; font-family: Verdana; font-size: 13.333333333333332px; vertical-align: baseline;">CACGTCTTTATAAGATTGAACACAATAAGGCAAATAGATT
 * CDS Gene Sequence (paste as text only):**


 * GC% Content for gene:**
 * 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 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):**