Klebsiella+Pneumoniae+Carbapenamase-2+(KPC-2)


 * Target: ** Klebsiella pneumoniae carbapenamase-2 (KPC-2)
 * *NCBI Gene # or RefSeq#: ** KP987218

//Klebsiella pneumoniae //is a bacterial species that frequently causes respiratory infections such as pneumonia but can also cause soft tissue infections in deadlier cases. Though //K. pneumoniae// is found in nature and can cause infection anywhere in nature, //K. pneumoniae// infection cases have increased steadily in hospitals, granting //K. pneumoniae// infections the title of “nosocomial infection” or hospital-originated infection [1]. Unfortunately, the reason for the increasing number of infections is due to the discovery of antibiotic resistance in recent strains of //K. pneumoniae// in the United States since 1996 [2]. Prior to 1996, beta-lactam drugs such as penicillin were sufficient to decrease infection. However, the evolution of beta-lactam resistant //K. pneumoniae// exacerbated the situation for infected patients. These new strains produce enzymes called beta-lactamases that hydrolyze beta-lactam antibiotics. Extremely broad spectrum antibiotics called carbapenems were synthesized and are used as a last-ditch effort to save a patient undergoing a deadly infection. Unfortunately, //K. pneumoniae// was discovered to have evolved to produce KPC-2, a carbapenamase that hydrolyzes carbapenems and leaves infected patients to suffer without any effective antibiotic [3].A solution to the antibiotic resistance of //K. pneumoniae// is to discover other chemical compounds that can inhibit carbapenamase and effectively prevent //K. pneumoniae// from being resistant to currently-available carbapenem antibiotics.
 * *Protein ID (NP or XP #) or Wolbachia#: **AHH27801
 * *Organism (including strain): **// Klebsiella pneumoniae //
 * Etiologic Risk Group: ** 2
 * */ Disease Information : **

**Is it a monomer or multimer as biological unit** ** ? ** Monomer
 * Link to TDR Targets page: ** None
 * Essentiality of this protein: ** Essential if KPC-2 is inactive with antibiotics in the environment of //K. pneumoniae//.
 * Complex of proteins?: **No.
 * Druggable Target: **// K. pneumoniae //

[]
 * *EC#: ** 3.5.2.6
 * Link to BRENDA EC# page: **http://www.brenda-enzymes.info/enzyme.php?ecno=3.5.2.6
 * Enzyme Assay information: **


 * Structure: ** PDB: 2OV5


 * Current Inhibitors: ** Clavulanic acid, tazobactam, penem 1 & 2, imipenem


 * Expression Information (has it been expressed in bacterial cells): ** Yes


 * Purification Method : ** Ni-NTA with 6xHis tag

Fig 2: X-ray crystal structure of KPC-2 (PDB: 2OV5). Active site shown as surface at 40% transparency; non-active site residues shown as cartoon; bicine molecule shown as sticks (carbons in cyan, oxygens in red, and nitrogens in blue). Protein coloration: carbons in green, oxygen in red, nitrogen in blue). Polar contacts shown as black dashes.
 * Image of protein (PyMol with features delineated and shown separately): **

MAEPFAKLEQDFGGSIGVYAMDTGSGATVSYRAEERFPLCSSFKGFLAAAVLARSQQQAGLLDTPIRYGKNALVPWSPISEKYL TTGMTVAELSAAAVQYSDNAAANLLLKELGGPAGLTAFMRSIGDTTFRLDRWELELNSAIPGDARDTSSPRAVTESLQKLTLGS ALAAPQRQQFVDWLKGNTTGNHRIRAAVPADWAVGDKTGTCGVYGTANDYAVVWPTGRAPIVLAVYTRAPNKDDKHSEAVIAAA ARLALEGLGV*
 * *Amino Acid Sequence: **


 * *length of your protein in Amino Acids: ** 263 aa


 * Molecular Weight of your protein in kiloDaltons: ** 27.768 kDa


 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 39420


 * TMpred graph Image: **

// ATG //// GCGGAACCATTCGCTAAAC //// TCGAACAGGAC // TTTGGCGGCTCCATCGGTGTGTACGCGATGGATACCGGCTCAGGCGCAACTGTAAGTTACCGCGCTGA GGAGCGCTTCCCACTGTGCAGCTCATTCAAGGGCTTTCTTGCTGCCGCTGTGCTGGCTCGCAGCCAGCAGCAGGCCGGCTTGCTGGACACACCCATCCGTT ACGGCAAAAATGCGCTGGTTCCGTGGTCACCCATCTCGGAAAAATATCTGACAACAGGCATGACGGTGGCGGAGCTGTCCGCGGCCGCCGTGCAATACAGT GATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCG CTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTG CACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGACCGGCAACCACCGCATCCGCGCGGCGGTGCCGGCAGACTGGGCAGTC GGAGACAAAACCGGAACCTGCGGAGTGTATGGCACGGCAAATGACTATGCCGTCGTCTGGCCCACTGGGCGCGCACCTATTGTGTTGGCCGTCTACACCCG GGCGCCTAACAAGGATGACAAGCACAGCGAGGCCGTCATCGCCGCTGCGGCTAGACTCGC GCTCGAGGGATTGGGCGTCTAACAGTAAAGGTGGATA
 * *CDS Gene Sequence **** : **


 * *GC% Content for gene: ** 61.3%


 * *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): **Primer overlap PCR was not done for KPC-2. Gene obtained from Fast lab in pET24-a vector.


 * *GC% Content for gene (codon optimized): **n/a


 * Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): ** n/a

Forward: 5’ TACTTCCAATCCATGGCGGAACCATTCGCTAAACT 3’ Reverse: 5’ TATCCACCTTTACTGTTAGACGCCCAATCCCTCGAG 3’