Difference between revisions of "Part:BBa K1045003"
 
(12 intermediate revisions by 2 users not shown)
Line 3: Line 3:
  
 
The severe threat caused by bacteria which are resistant to conventional antibiotic drugs and the appearance of even multi-resistant strains demonstrates the urgent need for the discovery of new antibacterial substance classes.
 
The severe threat caused by bacteria which are resistant to conventional antibiotic drugs and the appearance of even multi-resistant strains demonstrates the urgent need for the discovery of new antibacterial substance classes.
−
Cyclic di-AMP was discovered to be an essential signal molecule in Gram-positive bacteria including the pathogenic species ''Streptococcus pneumoniae'', ''Staphylococcus aureus'' and ''Listeria monocytogenes''. Both, loss and overproduction of c-di-AMP have detrimental effects on cell growth, cell wall synthesis, and propagation. Thus, the diadenylate cyclase (DAC) which catalyses the condensation reaction of two ATP molecules to c-di-AMP is the key factor for signal molecule production and maintenance of c-di-AMP homeostasis. We are convinced that the DAC is a very promising target for the development of highly specific antibiotic substances which exclusively act on Gram-positive bacteria and are not harmful to Gram-negative ones, including the gut bacterium ''Escherichia coli'' as well as humans. Here, we introduce a truncated but functional DAC which localizes to the cytosol and can easily be purified. Furthermore, protein crystals were obtained as well as the protein structure by X-ray diffraction analysis. The part [[Part:BBa_K1045003|BBa_K1045003]] described here, covers the amino acids 100 to 273 of the full length coding sequence of CdaA. <br /> For a precise description of this part, please vistit the experience page!
+
Cyclic di-AMP was discovered to be an essential signal molecule in Gram-positive bacteria including the pathogenic species ''Streptococcus pneumoniae'', ''Staphylococcus aureus'' and ''Listeria monocytogenes''. Both, loss and overproduction of c-di-AMP have detrimental effects on cell growth, cell wall synthesis, and propagation. Thus, the diadenylate cyclase (DAC) which catalyses the condensation reaction of two ATP molecules to c-di-AMP is the key factor for signal molecule production and maintenance of c-di-AMP homeostasis. We are convinced that the DAC is a very promising target for the development of highly specific antibiotic substances which exclusively act on Gram-positive bacteria and are not harmful to Gram-negative ones, including the gut bacterium ''Escherichia coli'' as well as humans. Here, we introduce a truncated but functional DAC which localizes to the cytosol and can easily be purified. <br />
 +
Furthermore, the protein was crystallized and the X-ray diffraction pattern obtained. This enabled the determination of the 3D-structure of DacA by Molecular Replacement using the similar diadenylate cyclase DisA from ''B. subtilis''.  
 
<p>
 
<p>
  
−
[[File:image002.png|400px|thumb|left|'''Fig. 1. c-di-AMP production and degradation.''' c-di-AMP is produced from two molecules of ATP by DACs and degraded to pApA by phosphodiasterases. (Edited from Corrigan and Gründling, 2013)]]
+
[[File:image002.png|370px|thumb|left|'''Fig. 1. c-di-AMP production and degradation.''' c-di-AMP is produced from two molecules of ATP by DACs and degraded to pApA by phosphodiasterases. (Edited from Corrigan and Gründling, 2013)]]
 
<html>
 
<html>
 
<object width="420" height="315"><param name="movie" value="//www.youtube.com/v/T0xpaOZtjfk?hl=zh_CN&amp;version=3&amp;rel=0"></param>
 
<object width="420" height="315"><param name="movie" value="//www.youtube.com/v/T0xpaOZtjfk?hl=zh_CN&amp;version=3&amp;rel=0"></param>
Line 13: Line 14:
 
<embed src="//www.youtube.com/v/T0xpaOZtjfk?hl=zh_CN&amp;version=3&amp;rel=0" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true">
 
<embed src="//www.youtube.com/v/T0xpaOZtjfk?hl=zh_CN&amp;version=3&amp;rel=0" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true">
 
</embed></object></html>
 
</embed></object></html>
−
</p>
+
</br> </p>
  
 +
The part [[Part:BBa_K1045003|BBa_K1045003]] described here, covers the amino acids 100 to 273 of the full length coding sequence of CdaA. For a precise description of this part, please vistit the experience page! <br />
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 15:25, 29 September 2018

Diadenylate cyclase domain of Listeria monocytogenes cdaA (DacA)

The severe threat caused by bacteria which are resistant to conventional antibiotic drugs and the appearance of even multi-resistant strains demonstrates the urgent need for the discovery of new antibacterial substance classes. Cyclic di-AMP was discovered to be an essential signal molecule in Gram-positive bacteria including the pathogenic species Streptococcus pneumoniae, Staphylococcus aureus and Listeria monocytogenes. Both, loss and overproduction of c-di-AMP have detrimental effects on cell growth, cell wall synthesis, and propagation. Thus, the diadenylate cyclase (DAC) which catalyses the condensation reaction of two ATP molecules to c-di-AMP is the key factor for signal molecule production and maintenance of c-di-AMP homeostasis. We are convinced that the DAC is a very promising target for the development of highly specific antibiotic substances which exclusively act on Gram-positive bacteria and are not harmful to Gram-negative ones, including the gut bacterium Escherichia coli as well as humans. Here, we introduce a truncated but functional DAC which localizes to the cytosol and can easily be purified.
Furthermore, the protein was crystallized and the X-ray diffraction pattern obtained. This enabled the determination of the 3D-structure of DacA by Molecular Replacement using the similar diadenylate cyclase DisA from B. subtilis.

Fig. 1. c-di-AMP production and degradation. c-di-AMP is produced from two molecules of ATP by DACs and degraded to pApA by phosphodiasterases. (Edited from Corrigan and Gründling, 2013)

</br>

The part BBa_K1045003 described here, covers the amino acids 100 to 273 of the full length coding sequence of CdaA. For a precise description of this part, please vistit the experience page!

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]