Proteomic Pipelines for the Quantification of Abundance and Turnover of Post-Translationally Modified Proteins in Aging Studies

National Institute On Aging

Investigators

Linked publications, trials & patents

Abstract

Building on our published studies that produced a novel computational workflow to accurately assess protein turnover rates in vivo in mice, we are now developing unique computational workflows to do the same in human studies. Equivalent human studies have unique technical challenges that are associated with metabolic labeling with heavy water. In ongoing work, we are developing a computational pipeline for human studies. Moreover, together with Dr. Ferrucci and the BLSA, we are conducting a human clinical study to assess in vivo protein turnover in humans across the lifespan in muscle, skin, fat, and blood. While the study is ongoing, we have collected several tissues and used them to show that we can successfully assess the half-lives of many proteins. Toward our goals of developing and applying workflows to identify proteoforms and PTMs, we have successfully applied a top-down discovery proteomics workflow to identifying proteoforms secreted by senescent cells for the first time. We will build on this by developing workflows to explore proteoforms in human blood samples. To explore the role of acetylation, a key PTM, in aging and health, we are conducting ongoing studies to enrich the acetylome at the peptide level and identify broad changes in the acetylome in aging skeletal muscle, both in untreated mice and mice given a supplement to boost NAD levels. In published collaborative work, we also identified proteomic changes in extracellular matrix, a key tissue that accumulates PTMs and modifications in the context of aging and cancer.