2023-2024 Faculty Research Grant

Peter Lyons (Biology).

Carboxypeptidase as a model for understanding protein folding.

Proteins are biological machines that are essential for all aspects of life. The three-dimensional structure of a protein is formed as the linear chain of its building blocks, the amino acids, fold together. However, the folding process often requires help from chaperones. Carboxypeptidase enzymes are proteins thought to require a particular piece (the prodomain) as an intramolecular chaperone to assist in folding. Our recent results suggest that a prodomain is unnecessary if specific amino acid changes are made to stabilize the end of the protein. We would like to further examine these results using a direct analysis of protein folding called a thermal shift assay, in which the rate of unfolding of a protein is seen as an increase in fluorescence of the SyPro Orange dye. This requires us to genetically engineer nucleic acid sequences so that they encode proteins that we can purify via metal affinity chromatography. In addition, we would like to extend these results to two other carboxypeptidase enzymes, to provide broader support for this hypothesis. These experiments will result in a better understanding of protein folding and misfolding, the cause of a variety of human diseases.