Enterotoxin+C3+(SEC+3), Staphylococcus+aureus

Format for Individual Target pages:
[|http://www.bacterio.net/-hazard.html#group2]
 * *Target (protein/gene name): **3-hydroxy-3-methylglutaryl coenzyme A reductase
 * *NCBI Gene # or RefSeq#: **NC_003098.1
 * *Protein ID (NP or XP #) or Wolbachia#: **A0A0D6J7E8
 * *Organism (including strain): **//Streptococcus pneumoniae R6 //
 * Etiologic Risk Group (see link below): ** Group 2


 * */ Disease Information (sort of like the Intro to your Mini Research Write up): **Pneumonia is an infection that affects the lungs and can result in mild or severe illness. There are various causes of pneumonia; viruses, bacteria, or fungi. //Streptococcus pneumoniae (pneumococcus) //is the bacterial cause of pneumonia, and 3-hydroxy-3-methylglutaryl coenzyme A reductase is a protein involved in the synthesis of (R)- mevalonate from (S)-3-hydroxy-3-methylglutaryl coenzyme A reductase. The mevalonate pathway is an essential metabolic pathway in eukaryotes and bacteria, and is responsible for the production of isoprenoids such as, cholesterol, heme, vitamin K, and steroid hormones.

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[|https://www.ncbi.nlm.nih.gov/gene/932918#reference-sequences] []
 * Link to TDR Targets page (if present): ** n/a
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **


 * Essentiality of this protein: ** The enzyme HMG-CoA reductase is an important factor in the mevolanate pathway which is found in various bacteria, eukaryotes, and archaea. The function of the mevalonate pathway is the production of isoprenoids such as, cholesterol, heme, vitamin K, and steroid hormones. Therefore, this pathway is often inhibited to reduce the production of cholesterol, similarly the inhibition of this pathway can eliminate the production of isopernoids produced by the //Streptococcus pneumoniae R6 //bacteria and kill it. HMG-CoA reductase is a key factor in the mevolante pathway as it synthesizes (R)- mevolante, inhibiting this enzyme would break the pathway and help reduce the effects of the bacteria or oil it altogether. Certain bacteria contain another pathway for isopernoids creation such as E. coli and //Mycobacterium tuberculosis//, however this is not found in //Streptococcus pneumoniae// so it would be effective.

[] Is it a monomer or multimer as biological unit ** ? (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): Multimer 3-hydroxy-3-methylglutaryl coenzyme A reductase is a draggable target as research for over expression, purification, and crystallography have already been completed for this target for antibiotic production. The prevalence of //Streptococcus pneumoniae R6 //necessitates various drug options due to antibiotic resistances.
 * Complex of proteins?: ** Yes
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **

h[|ttps://www.ncbi.nlm.nih.gov/pubmed/21045306/]

[|http://www.brenda-enzymes.org/enzyme.php?ecno=1.1.1.88&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Pseudomonas+mevalonii#EC%20NUMBER]
 * *EC#: 1.1.1.88 **
 * Link to BRENDA EC# page: **
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): ** X ray crystallography

[] Substrate information can be found on BRENDA
 * -- link to Sigma (or other company ) page for assay (see Sigma links below) **
 * -- -or link (or citation) to paper that contains assay information **
 * -- links to assay reagents (substrates) pages. **
 * --- List cost and quantity of substrate reagents, supplier, and catalog #: **

[|http://www.brenda-enzymes.org/enzyme.php?ecno=1.1.1.88&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Pseudomonas+mevalonii#SYSTEMATIC%20NAME]

-- PDB # or closest PDB entry if using homology model: 5WPK -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB Query Coverage: 100% Max % Identities: 99% % Positives: 99% Chain used for homology: A
 * Structure (PDB or Homology model) **


 * Current Inhibitors: **Multiple inhibitors listed on BRENDA
 * Expression Information (has it been expressed in bacterial cells): ** E. coli BL21 (DE3)
 * Purification Method : **Ni-NTA affinity chromatography

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code format="sequence"        10         20         30         40         50 MKISWNGFSK KSYQERLELL KAQALLSPER QASLEKDEQM SVTVADQLSE 60        70         80         90        100 NVVGTFSLPY SLVPEVLVNG QEYTVPYVTE EPSVVAAASY ASKIIKRAGG 110       120        130        140        150 FTAQVHQRQM IGQVALYQVA NPKLAQEKIA SKKAELLELA NQAYPSIVKR 160       170        180        190        200 GGGARDLHVE QIKGEPDFLV VYIHVDTQEA MGANMLNTML EALKPVLEEL 210       220        230        240        250 SQGQSLMGIL SNYATDSLVT ASCRIAFRYL SRQKDQGREI AEKIALASQF 260       270        280        290        300 AQADPYRAAT HNKGIFNGID AILIATGNDW RAIEAGAHAF ASRDGRYQGL 310       320        330        340        350 SCWTLDLERE ELVGEMTLPM PVATKGGSIG LNPRVALSHD LLGNPSAREL 360       370        380        390        400 AQIIVSIGLA QNFAALKALV STGIQQGHMK LQAKSLALLA GASESEVAPL 410       420 VERLISDKTF NLETAQRYLE NLRS code
 * 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 **426 amino acids
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: ** 46.294 kDa
 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 35995


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

code ATGAAGATAAGTTGGAATGGATTTTCTAAAAAATCATACCAAGAGCGCCTCGAGCTGCTAAAAGCTCAGG CGCTCCTTAGTCCTGAGAGACAAGCTAGTCTGGAGAAGGATGAACAGATGAGTGTGACTGTGGCAGACCA GCTGAGTGAGAATGTGGTGGGAACTTTTTCTCTGCCTTATTCGCTGGTTCCGGAGGTACTTGTCAACGGT CAGGAATACACCGTTCCCTATGTGACAGAAGAACCCTCTGTGGTTGCGGCGGCCAGCTATGCCAGCAAAA TCATCAAGCGTGCAGGTGGTTTTACTGCACAAGTCCATCAGCGCCAGATGATTGGGCAGGTAGCCCTTTA TCAAGTTGCTAATCCTAAACTAGCGCAAGAGAAGATTGCCAGCAAGAAAGCGGAGCTCTTGGAGCTTGCC AATCAAGCCTATCCTTCTATCGTTAAACGTGGGGGTGGGGCGCGTGATCTGCATGTCGAGCAGATAAAAG GCGAACCAGACTTTCTCGTTGTTTATATTCATGTCGATACCCAGGAAGCCATGGGTGCCAATATGCTCAA CACCATGCTGGAAGCCTTGAAACCAGTCTTAGAAGAACTCAGTCAGGGACAGAGTCTCATGGGAATCCTG TCCAACTACGCGACTGATTCTCTGGTGACTGCAAGCTGTCGCATCGCCTTTCGCTACTTGAGCCGCCAAA AGGATCAAGGACGAGAGATTGCGGAGAAAATTGCGTTGGCTAGTCAGTTTGCGCAGGCTGATCCTTACCG AGCTGCTACTCATAATAAAGGAATTTTTAATGGTATTGATGCGATTTTGATTGCCACTGGTAATGACTGG CGTGCCATCGAAGCTGGGGCCCATGCCTTTGCCAGTCGAGATGGACGCTATCAAGGTCTTAGCTGCTGGA CGCTGGACCTTGAAAGAGAAGAATTGGTCGGTGAGATGACCCTGCCCATGCCTGTAGCGACTAAGGGTGG CTCTATCGGCCTCAACCCACGTGTAGCTCTCAGTCATGATCTACTAGGAAATCCTTCTGCCAGAGAATTA GCCCAGATTATCGTGTCCATCGGTCTTGCTCAAAATTTTGCAGCCCTCAAAGCCTTGGTAAGTACGGGCA TCCAGCAAGGCCACATGAAACTACAGGCCAAATCCCTAGCTCTCCTAGCTGGGGCTAGTGAATCTGAAGT TGCTCCCCTAGTAGAGCGCCTCATCTCAGATAAAACCTTTAACCTAGAGACAGCCCAGCGCTATCTCGAA AATTTAAGATCATAA code
 * *CDS Gene Sequence (paste as text only): **
 * *GC% Content for gene: ** G- 326, C- 303, GC%- 49.33%
 * *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): ** n/a
 * *GC% Content for gene (codon optimized): **n/a

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): **