CAREER: Engineering a Protein Molecular Switch

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The objective of this proposed research is to develop and implement a novel, combinatorial protein engineering strategy called random domain insertion to couple two proteins functionally to create a protein molecular switch. The design of molecular switches to modulate or report on biological functions has enormous potential for a variety of applications including the creation of biosensors, regulators of gene transcription, modulators of metabolic and cell signaling pathways, chaperones for drug delivery, facilitators of drug transport, and novel biomaterials. The efficacy of the random domain insertion strategy will be demonstrated in two models, "proof-of-principle" systems using well-characterized proteins. The kinetic, structural, and fluorescent behavior of representative molecular switches will be characterized so as to develop models on how the switching behavior was achieved. These models will be tested experimentally. The combinatorial methods and models developed will be utilized to initiate engineering a molecular switch for drug transport. The scientific insights gained from this will provide a rational basis for engineering switches for a variety of applications as well as provide a unique platform for addressing questions of structure-function relationships. For the education component, the PI will be developing an integrated program for infusing the Chemical Engineering Undergraduate Program at Johns Hopkins University with the proper components to prepare graduates who have a basic understanding of biology and who can apply quantitative and non-quantitative approaches to biological problems. This will include the development of a Biological Engineering Laboratory course.