Target-+phospholipase+C+(C.+Perfrigens)


 * *Target (protein/gene name): **phospholipase C


 * *NCBI Gene # or RefSeq#: ** NCTC8237

[]
 * *Protein ID (NP or XP #) or Wolbachia#: **GenBank: CAA31943.1

>gi|194293816|gb|EU839779.1| Clostridium perfringens strain S01 phospholipase C (plc) gene, complete cds


 * *Organism: ****Clostridium Perfrigens **


 * *Background/Disease Information: **//C. perfringens// is a bacteria that leads to necrosis, bacteremia and gas gangrene. It is the most common bacterial reagent leading to gas gangrene. It is the third most common cause of food poisoning in the United Kingdom and the United States.

**Molecular cloning and nucleotide sequence of the alpha-toxin (phospholipase C) of Clostridium perfringens. ** []
 * ESSENTIALITY**


 * Complex of enzymes: **


 * *EC#: **** 3.1.4.3 **


 * Link to BRENDA EC# page: **[]


 * -- ** Show screenshot of BRENDA enzyme mechanism schematic

[]
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **


 * -- **


 * Structure Available (PDB or Homology model) **


 * -- PDB # or closest PDB entry if using homology model: ****<span style="font-family: 'Calibri','sans-serif';">2WXT **


 * <span style="font-family: 'Calibri','sans-serif';">Druggable Target (see Databases for this): **


 * <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Inhibitors **
 * || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Inhibitors || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Commentary || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Organism || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Structure ||
 * <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">EDTA || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">enzyme activity against 1-myristoyl-lysophosphatidylcholine, glycerophosphorylcholine, and p-nitrophenyl phosphorylcholine are readily inhibited by EDTA. Indicates that a divalent cation is essential for catalytic activity || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Homo sapiens || <span style="font-family: 'Times New Roman','serif'; font-size: 16px; text-decoration: none;">[[image:http://vdsstream.wikispaces.com/site/embedthumbnail/placeholder?w=200&h=50 width="200" height="50" caption="Description: http://brenda-enzymes.org/images/structure.gif"]] ||
 * <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">EGTA || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">enzyme activity against 1-myristoyl-lysophosphatidylcholine, glycerophosphorylcholine, and p-nitrophenyl phosphorylcholine are readily inhibited by EGTA. Indicates that a divalent cation is essential for catalytic activity || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Homo sapiens || <span style="font-family: 'Times New Roman','serif'; font-size: 16px; text-decoration: none;">[[image:http://vdsstream.wikispaces.com/site/embedthumbnail/placeholder?w=200&h=50 width="200" height="50" caption="Description: http://brenda-enzymes.org/images/structure.gif"]] ||
 * <span style="color: #669933; font-family: 'Times New Roman','serif'; font-size: 16px;">additional information || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">the catalytic activity of NPP6 toward p-nitrophenyl phosphorylcholine is partially inhibited by the divalent cations. Attempts are unsuccessful to recover the activity of EDTA treated NPP6 by adding various divalent cations || <span style="font-family: 'Times New Roman','serif'; font-size: 16px;">Homo sapiens || <span style="display: block; font-family: 'Times New Roman',serif; font-size: 16px; text-align: center;">- ||  ||


 * <span style="font-family: 'Calibri','sans-serif';">Expression Information (has it been expressed in bacterial cells): **homo sapiens, E. Coli and mus musculus. Yes it was expressed in E. Coli

mono Q ion exchange chromatography column and eluted with a linear gradient of NaCl (0-2 M) using the deltaKTA system in homo sapiens and mus musculus
 * <span style="font-family: 'Calibri','sans-serif';">Purification Method: **



1 MSRLVVVSNRIAPPDEHAASAGGLAVGILGALKAAGGLWFGWSGETGNEDQPLKKVKKGN
 * *Amino Acid Sequence: ****<span style="font-family: Arial,sans-serif; font-size: 13.5pt;">* **

61 ITWASFNLSEQDLDEYYNQFSNAVLWPAFHYRLDLVQFQRPAWDGYLRVNALLADKLLPL

121 LQDDDIIWIHDYHLLPFAHELRKRGVNNRIGFFLHIPFPTPEIFNALPTYDTLLEQLCDY

181 DLLGFQTENDRLAFLDCLSNLTRVTTRSAKSHTAWGKAFRTEVYPIGIEPKEIAKQAAGP

241 LPPKLAQLKAELKNVQNIFSVERLDYSKGLPERFLAYEALLEKYPQHHGKIRYTQIAPTS

301 RGDVQAYQDIRHQLENEAGRINGKYGQLGWTPLYYLNQHFDRKLLMKIFRYSDVGLVTPL

361 RDGMNLVAKEYVAAQDPANPGVLVLSQFAGAANELTSALIVNPYDRDEVAAALDRALTMS

421 LAERISRHAEMLDVIVKNDINHWQECFISDLKQIVPRSAESQQRDKVATFPKLA

<span style="font-family: 'Times New Roman',serif; font-size: 12pt;">Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water 77935
 * *length of your protein in Amino Acids **<span style="font-family: Arial,sans-serif; font-size: 13.5pt;">474
 * <span style="font-family: 'Times New Roman',serif; font-size: 12pt;">Molecular Weight of your protein **<span style="font-family: 'Times New Roman',serif; font-size: 12pt;">53611.1


 * *GC% Content for gene: 34% **


 * Primer design **

1 ATGTCTCGTCTCGTTGTTGTTTCTAATCGTATCGCGCCTCCAGACGAACACGCGGCGTC 59

2 GCCGCCTTGAGGGCACCGAGGATACCCACCGCGAGGCCACCAGCAGACGCCGCGTGTTCG 60

3 TGCCCTCAAGGCGGCAGGCGGTCTGTGGTTCGGTTGGTCCGGTGAGACCGGTAACGAGGA 60

4 CCCAGGTGATGTTACCTTTCTTAACTTTTTTGAGTGGCTGATCCTCGTTACCGGTCTCAC 60

5 AGAAAGGTAACATCACCTGGGCGTCTTTCAACCTGTCTGAACAAGACCTGGACGAATACT 60

6 GGAACGCAGGCCAGAGAACGGCGTTAGAGAACTGGTTGTAGTATTCGTCCAGGTCTTGTT 60

7 TCTCTGGCCTGCGTTCCATTACCGTCTCGACCTGGTGCAATTTCAGCGTCCAGCGTGGGA 60

8 GGGAGGAGTTTATCCGCCAGCAGCGCGTTAACACGCAGATAGCCGTCCCACGCTGGACGC 60

9 TGGCGGATAAACTCCTCCCGCTCCTCCAGGACGACGATATCATCTGGATTCACGACTACC 60

10 CGCCACGCTTACGCAGCTCGTGCGCGAACGGCAGGAGGTGGTAGTCGTGAATCCAGATGA 60

11 CTGCGTAAGCGTGGCGTAAATAACCGTATCGGTTTCTTCCTGCACATCCCGTTCCCAACC 60

12 CAGCAGGGTGTCGTAGGTCGGGAGTGCGTTAAAGATTTCCGGGGTTGGGAACGGGATGTG 60

13 ACCTACGACACCCTGCTGGAACAGCTCTGTGACTACGACCTGCTCGGCTTCCAGACCGAG 60

14 GGGTCAGATTAGAGAGGCAGTCCAGAAACGCCAGACGGTCATTCTCGGTCTGGAAGCCGA 60

15 CTGCCTCTCTAATCTGACCCGTGTTACCACTCGTAGCGCGAAATCTCATACTGCGTGGGG 60

16 TTCGATACCGATAGGGTAAACTTCGGTGCGGAACGCTTTACCCCACGCAGTATGAGATTT 60

17 AAGTTTACCCTATCGGTATCGAACCGAAAGAAATCGCGAAACAAGCAGCCGGTCCGCTGC 60

18 TTCTGAACGTTTTTCAGTTCCGCTTTGAGCTGGGCCAGCTTTGGTGGCAGCGGACCGGCT 60

19 CGGAACTGAAAAACGTTCAGAACATCTTCTCTGTTGAACGCCTGGACTACTCTAAAGGTC 60

20 TTTCCAGGAGCGCTTCGTACGCCAGGAAACGCTCCGGCAGACCTTTAGAGTAGTCCAGGC 60

21 ACGAAGCGCTCCTGGAAAAATACCCGCAACATCATGGTAAAATTCGTTACACCCAGATTG 60

22 CTTGGTACGCCTGCACGTCGCCACGAGAGGTCGGCGCAATCTGGGTGTAACGAATTTTAC 60

23 CGTGCAGGCGTACCAAGACATCCGTCACCAACTGGAGAACGAAGCAGGCCGCATCAACGG 60

24 GGTTCAGGTAGTACAGCGGAGTCCAACCCAGCTGACCGTACTTACCGTTGATGCGGCCTG 60

25 CCGCTGTACTACCTGAACCAACACTTCGACCGTAAACTGCTGATGAAAATCTTCCGTTAC 60

26 TACCGTCGCGCAGTGGGGTCACCAGACCAACGTCAGAGTAACGGAAGATTTTCATCAGCA 60