# Structural profiling of native proteins using fluorosequencing, a single molecule protein sequencing technology

> **NIH NIH R43** · ERISYON INC · 2022 · $216,909

## Abstract

ABSTRACT
Despite the wealth of accurate and high sensitive data from next-gen DNA sequencing technologies,
understanding the impact of the observed genomic mutations or alterations in the transcript (such as from
splicing or transcriptional errors) on the protein structure and function is often unclear. To complicate this
understanding are observations, where small abundant proteoforms can cause toxic intracellular
aggregates. This link between genome/transcriptome to protein structure is unclear, because methods for
characterizing protein structures often involve recombinant protein production and the use of analytical
techniques that perform ensemble analysis. In this proposal, we introduce Fluorescent Protein Painting, a
workflow for obtaining protein’s structural profiles by expanding the Erisyon’s existing fluorosequencing
(single molecule protein sequencing) technology. The steps in workflow are - (a) reversible, covalent
capture of natively folded proteins, (b) fluorescently labeling select surface exposed residues, (c)
releasing, denaturing and digesting the proteins before (d) reacting the internal residues amino acid
residues with second set of fluorophores. Performing fluorosequencing on this population of peptides will
result in generation of protein structural profile or foot-prints on millions of individual peptides. Since the
proteins are directly obtained from their native environment and the method can discriminate different
protein conformations at single molecule sensitivity, we will have a better understanding of the impact of
genomic mutations on the functional state of the biological sample.

## Key facts

- **NIH application ID:** 10499925
- **Project number:** 1R43HG012561-01
- **Recipient organization:** ERISYON INC
- **Principal Investigator:** Jagannath Swaminathan
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $216,909
- **Award type:** 1
- **Project period:** 2022-09-22 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10499925

## Citation

> US National Institutes of Health, RePORTER application 10499925, Structural profiling of native proteins using fluorosequencing, a single molecule protein sequencing technology (1R43HG012561-01). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10499925. Licensed CC0.

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