Utilizing Unnatural Amino Acids To Illustrate Protein Structure–Function Relationships: An Experiment Designed for an Undergraduate Biochemistry Laboratory

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• 作者：Johnathan C. Maza ; Jordan K. Villa ; Lisa M. Landino ; Douglas D. Young
• 刊名：Journal of Chemical Education
• 出版年：2016
• 出版时间：April 12, 2016
• 年：2016
• 卷：93
• 期：4
• 页码：767-771
• 全文大小：227K
• 年卷期：0
• ISSN：1938-1328

文摘

The site-specific introduction of unnatural amino acids (UAAs) has been demonstrated to be a useful tool in protein engineering. Moreover, the incorporation of a UAA into a protein has become feasible with the increased commercial availability of UAAs and robust expression plasmids. In addition to the ease of incorporation, the concepts utilized in the incorporation of a UAA are ideal to illustrate key concepts involved in protein translation and structure–function relationships. Consequently, a novel experiment has been developed for the incorporation of various UAAs into the GFP fluorophore to alter spectral properties. This experiment provides students with a hands-on approach for transformation, protein expression and purification, and structure–function analysis.