# Proteomics and model organism humanization to decode human genetics

> **NIH NIH R35** · UNIVERSITY OF TEXAS AT AUSTIN · 2021 · $541,949

## Abstract

Summary/Abstract
 While the human genome provides a parts list of >20,000 proteins, it is still largely unknown how these
proteins assemble into `molecular machines' to carry out their biological roles. This is important both for basic
characterization of human genes and for understanding the mechanisms underlying most human genetic traits
and diseases, which often arise from defects in systems of proteins working together. We focus on the >5,000
human proteins shared across eukaryotes and dating to the last eukaryotic common ancestor. These ancient
proteins carry out critical cellular processes, including DNA replication, repair, transcription, splicing,
mitochondrial and ciliary processes, and trafficking, among others. They are disproportionately drivers of
human disease, linked to a wide array of disorders, spanning cancers, birth defects, metabolic disorders,
Parkinson disease, Huntington disease, amyotrophic lateral sclerosis, mental retardations, and more. More
than 750 of these deeply conserved human proteins are still entirely uncharacterized. A fundamental question
is how all of these proteins work together to support cell function. However, a key limitation remains the lack of
large-scale data directly interrogating these proteins' expression, interactions, and activation states. Current
approaches for quantifying the proteome are only beginning to survey the proteins expressed in mammalian
cells to any significant depth, and consistently suffer from low sensitivity and throughput. These limitations
have slowed medical applications, e.g. biomarker discovery, where techniques including mass spectrometry
and antibody arrays often lack sufficient sensitivity and quantification accuracy to be effective. We propose
research in three broad areas: First, we propose a major effort to biochemically define the main human protein
complexes, providing a mechanistic basis for interpreting diverse human genetics and diseases. We will focus
primarily on evolutionarily conserved human proteins, leveraging studies in other species, due to these
proteins' critical importance to cellular function. Second, we are developing surrogate functional assays for
deeply conserved human proteins by systematically humanizing yeast cells, replacing each essential yeast
gene in turn by its human version. The resulting strains serve as new physical reagents for studying human
genes in a simplified organismal context, opening up simple high-throughput assays of human gene function,
the impact of human genetic variation on gene function, the screening and repurposing of drugs, and the rapid
determination of mechanisms of drug resistance. Finally, we aim to advance a new proteomics technology,
single-molecule protein sequencing, which could potentially solve problems currently limiting the field, by
orders-of-magnitude improvements in sensitivity and throughput. Success of these aims will give new insights
into basic human cell biology and biochemistry, laying the founda...

## Key facts

- **NIH application ID:** 10085651
- **Project number:** 5R35GM122480-05
- **Recipient organization:** UNIVERSITY OF TEXAS AT AUSTIN
- **Principal Investigator:** EDWARD M MARCOTTE
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $541,949
- **Award type:** 5
- **Project period:** 2017-05-01 → 2022-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10085651

## Citation

> US National Institutes of Health, RePORTER application 10085651, Proteomics and model organism humanization to decode human genetics (5R35GM122480-05). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10085651. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*