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Development of a Site-Selective Protein Immobilization Methodology Utilizing Unnatural Amino Acids
Raliski, Benjamin K
Raliski, Benjamin K
Abstract
Proteins are nature’s catalysts and have evolved over millennia to be highly selective and efficient. As a result, many have sought to incorporate proteins into artificial systems to varying degrees of success. Immobilization of proteins onto solid supports can increase the stability of proteins in conditions that would normally induce denaturation but immobilization strategies can present their own challenges by using reactions that lack selectivity and can potentially disrupt protein function themselves. This work develops a new methodology for protein immobilization that uses an unnatural amino acid site-selectively incorporated into a protein as the functional handle for immobilization. As a model system, Green Fluorescent Protein was immobilized in this manner and found to retain its functional activity in conditions which would normally cause the protein to denature. Through the development of this methodology, it was found that immobilization efficiency is affected by the site where the functional handle is inserted, in addition to the accessibility and reactivity of the solid support. Finally, progress was made towards determining whether this methodology can be successfully incorporated into a system which utilizes microwave irradiation.
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Date
2015-05-01
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Chemistry