Protein degradation has two main methods: the more well-known lysosomal processes and the newly discovered non-lysosomal mechanisms. Our 2016 project, SWITCH, is focusing on a specific non-lysosomal mechanism called ClpXP. The system is composed of two subunits - ClpX, which linearizes the protein marked with a degradation tag, and ClpP, which degrades the protein into component amino acids. We have created an inducible genetic construct to regulate the transcription of ClpXP, giving the ability to characterize the degradation of chromoproteins; we have quantified the data using light reflection from the chromoprotein tsPurple using a prototype lightbox/camera. The further characterization of protein degradation is necessary for the precision and control necessary for the administration of medicines and the function of biosensors, specifically Georgia Tech’s Zinc biosensor.

The concentration of proteins in a cell is determined by both the amount synthesized and the amount naturally degraded. Thus, protein degradation is a crucial aspect of maintaining intramolecular equilibrium. A class of ATPases known as AAA+ Proteins contains a well-known proteolysis mechanism known as ClpXP in which the ClpX piece unfolds and translocates a tagged protein into a sequestered proteolytic compartment in ClpP. The tagged, linearized polypeptide is then degraded into free amino acids.