Difference between revisions of "Part:BBa K4229010"
 
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The SpyTag/Catcher system derives from the pathogen bacterium <i>Streptococcus pyogenes</i>. Thanks to the special chemical mechanism that this bacterium has developed, some of its proteins are highly stable. The surface compound SpyCatcher recognizes a small peptide chain of 13 amino acids, the SpyTag, spontaneously binding irreversibly to it through an isopeptide bond between a lysine (catcher) and an asparagine (tag) [1][2][3][4]. Since both SpyTag and SpyCatcher are made of canonical amino acids, they can be linked to proteins of interest of different organisms and can also be detected by antibodies. In addition, they bind to each other extremely specifically. This covalent bond is also very stable under different pH and temperature conditions [4]
  
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The SpyCatcher and SpyTag is a system, which was developed from from a surface protein from Streptococcus pyogenes, which makes it possible to link Proteins with each other. In our project, we linked the T1 protein of the wiffelballs with the spyTag to XiaI, of our indigo-pathway. XiaI is fused with the SpyCatcher as you can see in BioBrick BBa_K4229012.
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In our project, we fused the SpyCatcher to XiaI (BioBrick BBa_K4229012) for targeting to a microcompartment (wiffleball).
  
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<b>References</b>
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<br>
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[1] R. J. Dutton, D. Boyd, M. Berkmen, and J. Beckwith, “Bacterial species exhibit diversity in their mechanisms and capacity for protein disulfide bond formation” Proc. Natl. Acad. Sci. U. S. A., vol. 105, no. 33, pp. 11933–11938, 2008, doi: 10.1073/pnas.0804621105.<br>
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[2] D. Hatlem, T. Trunk, D. Linke, and J. C. Leo, “Catching a SPY: Using the SpyCatcher-SpyTag and related systems for labeling and localizing bacterial proteins” Int. J. Mol. Sci., vol. 20, no. 9, 2019, doi: 10.3390/ijms20092129.<br>
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[3] H. J. Kang and E. N. Baker, “Intramolecular isopeptide bonds: Protein crosslinks built for stress?” Trends Biochem. Sci., vol. 36, no. 4, pp. 229–237, 2011, doi: 10.1016/j.tibs.2010.09.007.<br>
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[4] B. Zakeri et al., “Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin” Proc. Natl. Acad. Sci. U. S. A., vol. 109, no. 12, 2012, doi: 10.1073/pnas.1115485109.
  
  

Latest revision as of 23:00, 11 October 2022


SpyCatcher


The SpyTag/Catcher system derives from the pathogen bacterium Streptococcus pyogenes. Thanks to the special chemical mechanism that this bacterium has developed, some of its proteins are highly stable. The surface compound SpyCatcher recognizes a small peptide chain of 13 amino acids, the SpyTag, spontaneously binding irreversibly to it through an isopeptide bond between a lysine (catcher) and an asparagine (tag) [1][2][3][4]. Since both SpyTag and SpyCatcher are made of canonical amino acids, they can be linked to proteins of interest of different organisms and can also be detected by antibodies. In addition, they bind to each other extremely specifically. This covalent bond is also very stable under different pH and temperature conditions [4]

In our project, we fused the SpyCatcher to XiaI (BioBrick BBa_K4229012) for targeting to a microcompartment (wiffleball).


References
[1] R. J. Dutton, D. Boyd, M. Berkmen, and J. Beckwith, “Bacterial species exhibit diversity in their mechanisms and capacity for protein disulfide bond formation” Proc. Natl. Acad. Sci. U. S. A., vol. 105, no. 33, pp. 11933–11938, 2008, doi: 10.1073/pnas.0804621105.
[2] D. Hatlem, T. Trunk, D. Linke, and J. C. Leo, “Catching a SPY: Using the SpyCatcher-SpyTag and related systems for labeling and localizing bacterial proteins” Int. J. Mol. Sci., vol. 20, no. 9, 2019, doi: 10.3390/ijms20092129.
[3] H. J. Kang and E. N. Baker, “Intramolecular isopeptide bonds: Protein crosslinks built for stress?” Trends Biochem. Sci., vol. 36, no. 4, pp. 229–237, 2011, doi: 10.1016/j.tibs.2010.09.007.
[4] B. Zakeri et al., “Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin” Proc. Natl. Acad. Sci. U. S. A., vol. 109, no. 12, 2012, doi: 10.1073/pnas.1115485109.


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]