Difference between revisions of "Team:GeorgiaTech/Description"

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<h2>Our Purpose</h2>
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<h2> What's the Big Deal About Click Chemistry?</h2>
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<p style="font-size:25px; font-weight:bold;">From the heart of Atlanta, GA, we, the <a style="font-size:25px"; href="https://2015.igem.org/Team:GeorgiaTech/Members">2015 Georgia Tech iGEM team</a>, enthusiastically work to make an impact on the scientific community. We ambitiously (yet respectfully) challenge a paradigm of enzyme biology in our pursuit to design a novel enzyme with unnatural functions. </p>
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The copper-catalyzed azide-alkyne cycloaddition (<a href =  
  
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<p style="font-size:25px">The goal of our project is to discover an enzyme that will aid catalysis of the copper-catalyzed azide-alkyne cycloaddition (<a href="https://2015.igem.org/Team:GeorgiaTech/Background"; style="font-size:25px">CuAAC</a>) reaction in conditions where free copper is scarce. We will create a diverse <a href="https://2015.igem.org/Team:GeorgiaTech/Background#library"; style="font-size:25px">library of mutant proteins</a> stemming from six naturally occurring <a href="https://2015.igem.org/Team:GeorgiaTech/Background#starting"; style="font-size:25px">copper-binding proteins</a>.</p>
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"https://2015.igem.org/Team:GeorgiaTech/Background">CuAAC</a>) reaction  
  
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<p>The CuAAC reaction is a powerful and exciting reaction that has the potential to greatly advance the field of <i>in vivo</i> organic synthesis because it is modular, versatile, and consistently provides product with little to no side product formation. The azide and alkyne that participate in this reaction invariably produce the thermodynamic product, a stable triazole heteroatom ring. Furthermore, this reaction’s reactivity is strictly controlled by the absence or presence of a copper catalyst, making it highly selective (Sharpless, Kolb, Finn, 2001). The unique combination thermodynamic and kinetic properties of the CuAAC reaction inspire us to find applications for it in delicate systems such as in living cells. The reaction is also bioorthogonal, meaning that it does not interfere with biological function in any way, so it has great potential for drug delivery systems with minimal side effects. The catalyst -- a copper ion in the +1 oxidation state -- is the key to triggering the reaction, but free copper is extremely scarce in cells due to its cytotoxicity (Biaglow, 2010). As a result, most of the copper found in cells (about 70 uM) is bound to select protein complexes that require copper (Rae, et al., 1999).</p>
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has been widely used in the laboratory for tagging or labeling
  
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<p>The CuAAC reaction does not occur in nature; there is not a natural enzyme that is designed to accelerate the reaction. According to our current understanding of protein promiscuity, designing an enzyme with novel functions has not been documented; the closest we can get is amplifying an enzyme’s residual function until this residual function is dominant over the enzyme’s original function. Furthermore, most natural copper-binding proteins suppress copper’s reactivity during cellular transport by burying it within the protein structure, and this sequestration is detrimental to our efforts (Huffman, 2001). This makes the task that we have assigned ourselves daunting, but we believe that the properties of certain natural copper-binding proteins show promise in their abilities to be evolved from their natural functions into proteins that can catalyze the CuAAC reaction. We have selected six copper-binding proteins that both bind copper very tightly (K<sub>d</sub> values around 10<sup>-18</sup>), and yet also exchange Cu(I) very rapidly. This dual function can be attributed to the rare exterior positioning of the copper-binding sites in these proteins. Since our selected proteins are strong copper binders and expose copper on their surfaces, we are optimistic that the proteins can be artificially evolved to accelerate the CuAAC reaction.</p>
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biological molecules (Hong, 2010), but it cannot be directly applied
  
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<p>Successful identification and characterization of an enzyme that binds to copper in order to catalyze the CuAAC reaction will not only allow us to further optimize a powerful reaction that could completely change the current approach to drug delivery--this new enzyme will overturn a decade-old paradigm.</p>
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in living organisms because of the toxicity associated with excess or
  
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free copper ions (Biaglow, 1997). The development of an enzyme to
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<h3>Bibliography</h3>
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<ol><li>Biaglow, J. E.; Manevich, Y.; Uckun, F.; Held, K. D. <i>Free Radic Biol Med</i> 1997, 22, 1129.</li>
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<li>Huffman, D. L.; O'Halloran, T. V. <i>Annu Rev Biochem</i> 2001, 70, 677.</li>
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<li>Kolb, H. C.; Finn, M. G.; Sharpless, K. B. <i>Angew. Chem. Int. Ed.</i> 2001, 40: 2004–2021.</li>
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<li>Rae, T. D.; Schmidt, P. J.; Pufahl, R. A.; Culotta, V. C.; O’Halloran, T. V. <i>Science</i> 1999, 284 (5415), 805-808.</li>
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catalyze the CuAAC reaction would perfectly complement the growing
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<h5>What should this page contain?</h5>
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<li> A clear and concise description of your project.</li>
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<li>A detailed explanation of why your team chose to work on this particular project.</li>
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<li>References and sources to document your research.</li>
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<li>Use illustrations and other visual resources to explain your project.</li>
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ability of scientists to introduce azide- and alkyne-labeled molecules
  
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into biological systems (Liu, 1999). Our goal is to discover a protein
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<h4>Advice on writing your Project Description</h4>
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to bind Cu ions safely <i>in vivo</i> and perform the CuAAC reaction
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We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.
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through an innovative approach. We will attempt this by <a href
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Judges like to read your wiki and know exactly what you have achieved. This is how you should think about these sections; from the point of view of the judge evaluating you at the end of the year.
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</p>
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="https://2015.igem.org/Team:GeorgiaTech/Background#library">generating
  
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a large library</a> of Cu-binding proteins. Later teams will develop a
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<h4>References</h4>
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<p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you though about your project and what works inspired you.</p>
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reliable <a href =
  
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"https://2015.igem.org/Team:GeorgiaTech/Background#phage">phage
  
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<h4>Inspiration</h4>
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display</a> system to screen for the desired CuAAC activity, and  
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<p>See how other teams have described and presented their projects: </p>
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evolving any active enzymes for improved function.
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<li><a href="https://2014.igem.org/Team:Imperial/Project"> Imperial</a></li>
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<li><a href="https://2014.igem.org/Team:UC_Davis/Project_Overview"> UC Davis</a></li>
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<li><a href="https://2014.igem.org/Team:SYSU-Software/Overview">SYSU Software</a></li>
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<h4>Bibliography</h4>
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<ol><li>Biaglow, J. E.; Manevich, Y.; Uckun, F.; Held, K. D. <i>Free
  
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Radic Biol Med</i> 1997, 22, 1129.</li>
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<li>Hong, V.; Steinmetz, N. F.; Manchester, M.; Finn, M. G.
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<i>Bioconjug Chem</i> 2010, 21, 1912.</li>
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<li>Liu, D. R.; Schultz, P. G. <i>Proc Natl Acad Sci U S A</i> 1999,
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96, 4780.</li></ol>
 
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Revision as of 21:03, 18 September 2015

What's the Big Deal About Click Chemistry?

The copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction has been widely used in the laboratory for tagging or labeling biological molecules (Hong, 2010), but it cannot be directly applied in living organisms because of the toxicity associated with excess or free copper ions (Biaglow, 1997). The development of an enzyme to catalyze the CuAAC reaction would perfectly complement the growing ability of scientists to introduce azide- and alkyne-labeled molecules into biological systems (Liu, 1999). Our goal is to discover a protein to bind Cu ions safely in vivo and perform the CuAAC reaction through an innovative approach. We will attempt this by generating a large library of Cu-binding proteins. Later teams will develop a reliable phage display system to screen for the desired CuAAC activity, and evolving any active enzymes for improved function.

Bibliography

  1. Biaglow, J. E.; Manevich, Y.; Uckun, F.; Held, K. D. Free Radic Biol Med 1997, 22, 1129.
  2. Hong, V.; Steinmetz, N. F.; Manchester, M.; Finn, M. G. Bioconjug Chem 2010, 21, 1912.
  3. Liu, D. R.; Schultz, P. G. Proc Natl Acad Sci U S A 1999, 96, 4780.