TargetSp17+-+Biotin+carboxylase+(Staphylococcus+aureus)


 * Target (protein/gene name): **Biotin Carboxylase
 * NCBI #: **Reference Sequence NC_002758.2
 * Protein ID (NP or XP #) or Wolbachia#: ** BAB57768
 * Organism (including strain): **Methicillin-resistant //Staphylococcus aureus, Mu50//
 * Etiologic Risk Group (see link below): ** RG2

MRSA is a bacterial infection from Methicillin-resistant //Staphylococcus aureus.// Traditionally//, S. aureus// causes skin infections (cellulitis, impetigo), food poisoning, and bacteremia. Recently, strains of //S. aureus// have become resistant to common beta-lactam antibiotics, which include penicillins. This resistance does not make MRSA more virulent, but does make it substantially more difficult to treat. MRSA is especially dangerous in hospitals, where those with weakened immune systems are more prone to infection. Worldwide, up to 53 million people are thought to carry MRSA. The current treatment of MRSA are glycopeptide antibiotics such as vancomycin, however there are strains of //S. aureus// that have become resistant even to vancomycin (VISA).
 * Disease Information (sort of like the Intro to your Mini Research Write up): **
 * Link to TDR Targets page (if present): ** n/a
 * Link to Gene Database page NCBI: **https://www.ncbi.nlm.nih.gov/protein/BAB57768.1

Bacterial acetyl-CoA carboxylase is an essential enzyme in bacteria because it catalyzes the first step in fatty acid synthesis. The enzyme produces malonyl-CoA, which is a building block for fatty acids. Fatty acids can then be incorporated into phospholipids, which form much of the lipid bilayer in the cell. Inhibition of this enzyme will deprive the bacteria of fatty acids, which can lead to cell death and inability to reproduce since the cell membrane cannot be replenished. Furthermore, products of this pathway are used in quorum sensing and protein modification. **Is it a monomer or multimer as biological unit:** dimer
 * Essentiality of this protein: **

Biotin carboxylase is part of bacterial acetyl-CoA carboxylase, an enzyme that contains three separate proteins: biotin carboxylase (BC), biotin carboxyl carrier protein (BCCD), and carboxyltransferase (CT). Prototype inhibitor found for biotin carboxylase in [|Haemophilus influenzae] A continuous, spectrophotometric assay is available. [|Link to paper.] Can we just measure the BC activity? - Maybe just use Biotin and not BCCP? - Dr. B 060617 BC only assay [|here], and [|another one]
 * Complex of proteins?: **
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **
 * *EC#: **6.3.4.14
 * Link to BRENDA EC# page: **http://www.brenda-enzymes.org/enzyme.php?ecno=6.3.4.14
 * Enzyme Assay information: **
 * -- links to assay reagents (substrates) pages. **
 * --- List cost and quantity of substrate reagents, supplier, and catalog # **

[|PDB] #: 2VPQ In eukaryotes, the BC, CT and BCCD subunits reside in a single polypeptide chain. In bacteria, ACC assembles as a multisubunit complex with each of the reactions performed by a separate enzyme. These differences in tertiary structure can provide for selective inhibitors.
 * Structure (PDB or Homology model) **
 * Similarity to Human:**

One weak inhibitor [|has been discovered]. Chain A Sequence: MKKVLIANRGEIAVRIIRACRDLGIQTVAIYSEGDKDALHTQIADEAYCVGPTLSKDSYLNIPNILSIATSTGCDGVHPGYGFLAENADFAELCEACQLKFIGPSYQSIQKMGIKDVAKAEMIKANVPVVPGSDGLMKDVSEAKKIAKKIGYPVIIKATGGGGKGIRVARDEKELETGFRMTEQEA QTAFGNGGLYMEKFIENFRHIEIQIVGDSYGNVIHLGERDCTIQRRMQKLVEEAPSPILDDETRREMGNAAVRAAKAVNYENAGTIEFIYDLNDNKFYFMEMNTRIQVEHPVTEMVTGIDLVKLQLQVAMGDVLPYKQEDIKLTGHAIEFRINAENPYKNFMPSPGKIEQYLAPGGYGVRIESA CYTNYTIPPYYDSMVAKLIIHEPTRDEAIMAGIRALSEFVVLGIDTTIPFHIKLLNNDIFRSGKFNTNFLEQNSIMNDEG Chain B Sequence: MKKVLIANRGEIAVRIIRACRDLGIQTVAIYSEGDKDALHTQIADEAYCVGPTLSKDSYLNIPNILSIATSTGCDGVHPGYGFLAENADFAELCEACQLKFIGPSYQSIQKMGIKDVAKAEMIKANVPVVPGSDGLMKDVSEAKKIAKKIGYPVIIKATAGGGGKGIRVARDEKELETGFRMTEQE AQTAFGNGGLYMEKFIENFRHIEIQIVGDSYGNVIHLGERDCTIQRRMQKLVEEAPSPILDDETRREMGNAAVRAAKAVNYENAGTIEFIYDLNDNKFYFMEMNTRIQVEHPVTEMVTGIDLVKLQLQVAMGDVLPYKQEDIKLTGHAIEFRINAENPYKNFMPSPGKIEQYLAPGGYGVRIES ACYTNYTIPPYYDSMVAKLIIHEPTRDEAIMAGIRALSEFVVLGIDTTIPFHIKLLNNDIFRSGKFNTNFLEQNSIMNDEG ATGAAAAAGGTTTTAATTGCAAACCGCGGTGAAATCGCAGTTAGGATTATTCGCGCTTGTCGTGATTTAGGCATCCAAACTGTTGCAATCTATTCTGAAGGGGATAAAGATGCGCTACATACTCAAATTGCTGATGAAGCATATTGCGTAGGTCCCACTTTGTCTAAAGATTCATATTTAAATATTCCGAACATCTTATCT ATTGCAACTTCTACAGGTTGTGATGGCGTCATCCGGGTTATGGCTTTTTAGCTGAAAATGCTGACTTTGCAGAATTATGCGAAGCATGCCAATTGAAGTTCATTGACCAAGTTATCAATCTATCCAAAAAATGGGTATCAAAGATGTTGCTAAGGCAGAAATGATCAAAGCCAATGTTCCAGTTGTTCCTGGTAGTGAC GGTTTAATGAAAGACGTCTCAGAAGCTAAGAAAATCGCTAAGAAAATTGGCTATCCGGTCATCATTAAAGCTACTGCTGGTGGTGGCGGAAAAGGTATCCGTGTTGCTCGTGATGAAAAAGAACTTGAAACTGGCTTCCGAATGACAGAACAAGAAGCTCAAACTGCATTTGGTAATGGTGGACTTTATATGGAGA AATTCATCGAAAACTTCCGCCATATTGAAATCCAAATTGTTGGGGACAGCTATGGTAATGTAATTCATTTAGGAGAACGTGATTGTACAATTCAAAGACGTATGCAGAAATTAGTGGAAGAAGCACCTTCCCCAATTTTAGATGATGAAACACGTCGTGAAATGGGAAATGCCGCAGTTCGTGCAGCGAAAGCTGTA AATTATGAAAATGCGGGAACAATTGAGTTTATATATGATTTAAATGATAATAAATTTTATTTTATGGAAATGAATACACGTATTCAAGTAGAACATCCTGTAACTGAAATGGTAACAGGAATTGATTTAGTTAAATTACAATTACAAGTTGCTATGGGTGACGTGTTACCGTATAAACAAGAAGATATTAAATAACAGGACA CGCAATTGAATTTAGAATTAATGCTGAAAATCCTTACAAGAACTTTATGCCATCACCAGGTAAAATTGAGCAATATCTTGCACCAGGTGGATATGGTGTTCGAATAGAGTCAGCATGTTATACTAATTATACGATACCGCCATATTATGATTCGATGGTAGCGAAATTAATCATACATGAACCGACACGAGATGAAGCGAT TATGGCTGGCATTCGTGCATTAAGTGAATTTGTGGTTCTTGGTATTGATACAACTATTCCATTCCATATTAAATTATTGAATAACGATATATTTAGAAGCGGTAAATTTAATACAAACTTTTTAGAGCAAAATAGCATTATGAATGATGAAGGTTAA
 * Current Inhibitors: **
 * Expression Information (has it been expressed in bacterial cells): **It can and has been expressed in //E. coli.// Expressed in cytoplasm. ACC is essential for growth, is expressed at all times, expression is also very regulated to maintain the correct stoichiometry of subunits.
 * Purification Method : ** Standard nickel affinity column chromatography can be used.
 * 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: ** 451 amino acids.
 * Molecular Weight of your protein in kiloDaltons: ** 50.05 kDA
 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 28685 mol^-1cm-1
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html).
 * *CDS Gene Sequence (paste as text only): **

Do Not Need this info for Spring (but still copy these lines to your Target page for now)


 * *GC% Content for gene: **


 * output**


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

-- 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): **