This project aims to enhance understanding of the aging process through the study of the long-lived Caenorhabditis elegans daf-2 mutant. A data-independent-acquisition mass spectrometry scheme will be developed to enable a high-sensitivity, high-dynamic range, quantifiable analysis of the C. elegans proteome throughout the aging process. To this end, software will be written to deconvolute MS/MS spectra derived from the fragmentation of multiple precursors. C. elegans, with a germ-line mutation to avoid development of offspring, will be synchronized and grown. Strains that will be grown include a glp-4 strain, a glp-4;daf-2 mutant strain, and a daf-16 glp-4;daf-2 mutant strain. The protein profile of these strains will be analyzed over time to uncover new components of the aging pathway as well as generate a metric for "molecular age". The relationship between "molecular age" and chronological age will be determined. PUBLIC HEALTH RELEVANCE: This project will provide insight into the aging process and possibly the relationship between caloric restriction and longevity which has been observed in humans. The project will attempt to uncover the mechanism that allows a species of nematode (Caenorhabditis elegans) to double its lifespan. An understanding of this mechanism could open the door to new methods of studying and slowing human aging.