# Systems-to-structure approaches for defining mitochondrial protein function

> **NIH NIH R35** · WASHINGTON UNIVERSITY · 2024 · $554,165

## Abstract

PROJECT SUMMARY
Mitochondria are centers of metabolism and signaling, and their functions are essential for all but a few eukaryotic
cell types. Despite the widespread importance of these organelles, many aspects of mitochondrial biology remain
remarkably obscure. This fact contributes to our poor ability to diagnose mitochondrial diseases and our near
complete inability to rectify mitochondrial dysfunction therapeutically. This dysfunction is associated with ~350
rare inborn errors of metabolism and an increasing number of common diseases—including Parkinson’s,
Alzheimer’s, various cancers, and type 2 diabetes—often through distinct, yet unclear means. A major bottleneck
to further understanding mitochondrial processes and addressing their dysfunction in human disease is that the
proteins driving them have often not been identified. Concurrently, the basic biochemical functions of many
mitochondrial proteins that may fulfill these roles are undefined, or at best are poorly understood. This reality
was made manifest by my efforts to generate MitoCarta, which doubled the number of known mammalian
mitochondrial proteins and revealed a striking ~300 lacking any annotated function. Thus, an overarching goal
of my research program is to achieve a more comprehensive understanding of mitochondrial biology by
systematically establishing the functions of orphan mitochondrial proteins and their roles within disease-related
processes. We do so by first devising novel, multi-dimensional analyses designed to make new connections
between these proteins and established pathways and processes. These include customized, multiomic
analyses of yeast and human cell gene knockouts, deep mutational scanning approaches, large-scale genetic
screens, and other mass spectrometry-based investigations. We then employ mechanistic and structural
approaches to define the functions of select proteins at biochemical depth, and chemical biology approaches to
design small molecules to manipulate their functions in vitro and in vivo. This strategy leads us into diverse and
unanticipated new directions; however, we also maintain a persistent focus on the biochemistry, biosynthesis,
and transport of coenzyme Q (CoQ). We are driven to define how this remarkable, redox-active, extremely
hydrophobic molecule is produced in mitochondria, distributed throughout the cell, and how its cofactor and
antioxidant functions empower an ever-growing list of diverse biochemical processes. Overall, these “systems
biochemistry” efforts promise to help establish a deep, mechanistic understanding of mitochondrial biochemistry
that will motivate novel therapeutic strategies for the vast array of human disorders rooted in mitochondrial
dysfunction.

## Key facts

- **NIH application ID:** 10842904
- **Project number:** 2R35GM131795-07
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** David J Pagliarini
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $554,165
- **Award type:** 2
- **Project period:** 2019-04-01 → 2029-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10842904

## Citation

> US National Institutes of Health, RePORTER application 10842904, Systems-to-structure approaches for defining mitochondrial protein function (2R35GM131795-07). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10842904. Licensed CC0.

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