# Chemoselective assembly of multiply-labeled proteins for single molecule applications

> **NIH NIH R21** · SCRIPPS RESEARCH INSTITUTE, THE · 2020 · $241,875

## Abstract

Abstract
Single molecule techniques have fueled key insights into the molecular basis of protein function by facilitating
the ability to characterize individual conformational populations and dynamics. In recent years, there has been
an explosion in the capabilities of instrumentation, and a vast number of biological questions have been
identified that could be addressed using this technique, especially if multiple distances could be simultaneously
measured in large protein molecules (requiring 3-6 site specific labels). However, methodologies for generating
proteins with the requisite labels have lagged, limiting applications needing multi-site labeling. We propose to
develop, adapt and combine a robust set of protein synthesis / semisynthesis methodologies centered around
the mechanistically related chemistries of native chemical ligation and protein trans splicing to the challenge of
developing novel tools for building multiply-labeled proteins. Through careful synthetic design, we have
developed a strategy that will facilitate site-specific protein labeling at multiple positions in a protein using a
single, commonly-practiced labeling reaction. This semisynthetic approach is compatible with any protein,
regardless of length or sequence and has the potential to close the gap between experiments that can be
imagined and those that can be performed in the laboratory.

## Key facts

- **NIH application ID:** 9937779
- **Project number:** 5R21GM132787-02
- **Recipient organization:** SCRIPPS RESEARCH INSTITUTE, THE
- **Principal Investigator:** PHILIP E DAWSON
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $241,875
- **Award type:** 5
- **Project period:** 2019-06-01 → 2022-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9937779

## Citation

> US National Institutes of Health, RePORTER application 9937779, Chemoselective assembly of multiply-labeled proteins for single molecule applications (5R21GM132787-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9937779. Licensed CC0.

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