ATPGobble: Tool to Study the Regulatory Roles of ATP/ADP Ratio in Living Organisms
Denis Titov, Assistant Adjunct Professor
Nutritional Sciences and Toxicology
Closed. This professor is continuing with Fall 2023 apprentices on this project; no new apprentices needed for Spring 2024.
Adenosine triphosphate (ATP) fuels most energy-requiring reactions in living organisms through its degradation into adenosine diphosphate (ADP) and phosphate. This fuel capacity relies on a high ATP/ADP ratio that exceeds the thermodynamic equilibrium. High ATP/ADP ratio is critical for cell survival, and its perturbations have been connected with disease and aging. The most robust interventions to extend the healthy lifespan -- caloric restriction and exercise -- involve a transient drop in ATP/ADP ratio with subsequent adjustment. However, the causal nature of this relationship is poorly understood, and many mechanisms connecting the challenge to ATP/ADP ratio with resulting adaptation and fitness remain unknown. We are using the bacterial F1 ATPase as what we call ATPGobble, a tool that decreases ATP/ADP ratio in vivo, helping study the effects of ATP/ADP ratio on cell physiology, health, and lifespan. Because it is genetically encoded, ATPGobble can be targeted to a desired organism, tissue, or even cell compartment, offering the level of specificity and flexibility that the conventional tools, such as drugs or knock-out models, do not have. The overarching aim of this project is to validate the use of ATPGobble as a research tool, as well as use it to answer questions such as: the effect of ATP/ADP ratio on cell growth and survival; the effect of ATP/ADP ratio on protein expression; the effect of ATP/ADP ratio on insulin secretion; the effect of ATP/ADP ratio on lifespan in C. elegans.
Role: The main role for the undergraduate would be generation and quality control of cell lines expressing ATPGobble, though the exact responsibilities are negotiable. Techniques available to learn include: every-day cell culture, transduction and transfection; protein work and western blotting; cell staining and different kinds of fluorescence microscopy; subcellular fractionation; Seahorse (common instrument for metabolic analysis). Our lab has weekly lab meetings where the student will be able to learn about the biochemistry of metabolic enzymes, mathematical modeling of metabolism, and the use of C. elegans as a model of metabolism and aging (attendance optional).
All contributions to the advancement of the project will lead to inclusion as an author on future presentations and publications. As an estimate, this project is likely to be submitted for publication within 1 to 2 years, and the placement could be continued for up to 2 years.
Qualifications: 1) On-going studies in molecular and cell biology (or similar) with interest in metabolism and aging. 2) Being organized and eager to learn, such as: punctuality; ability to document work and be transparent about any mistakes or problems; genuine interest in the project. 3) Once the offer is extended and accepted, no concurrent apprenticeships.
Day-to-day supervisor for this project: Alex Ekvik, Ph.D. candidate
Hours: 12 or more hours
Related website: http://
Biological & Health Sciences