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

• Johnathan C. Maza ^[1] ; Jordan K. Villa ^[1] ; Lisa M. Landino ^[1] ; Douglas D. Young ^[1]
  1. [1] College of William and Mary, United States
• Localización: Journal of chemical education, ISSN 0021-9584, Vol. 93, Nº 4, 2016, págs. 767-771
• Idioma: inglés
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• Resumen

  • 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.