# Chemical Approaches to Understanding Reversible Lysine Modifications

> **NIH NIH R35** · BRIGHAM AND WOMEN'S HOSPITAL · 2024 · $447,500

## Abstract

Abstract
This is an R35 NIGMS application that is meant to succeed R37GM62437 and concerns the development and
application of chemical approaches to enhance our understanding of histone post-translational modifications
(PTMs) on Lys residues and the enzymes that attach and remove them (“writers” and “erasers”). Our lab has a
record of technical innovation in the chemical biology of histone modifications including bivalent analog design
and protein semisynthesis. Our bivalent compound approaches have led to the generation of potent and
selective inhibitors of p300 and CBP (p300/CBP) acetyltransferases and the LSD1-HDAC1-CoREST (LHC)
demethylase/ deacetylase gene silencing complex. Our latest-generation compounds, A485 for p300/CBP and
corin for LHC have become very useful and popular pharmacological probes for analyzing these histone-
modifying enzymes in mechanistic and preclinical therapeutic experiments. Our protein semisynthetic methods
including expressed protein ligation, engineered sortase-catalyzed histone production, and Cys modification to
introduce acyl-Lys mimics have proven to be efficient approaches to furnish site-specifically modified proteins.
In recent years we have shown how particular histone modifications influence nucleosome stability and
susceptibility to eraser enzymes including deacetylase, demethylase, and deubiquitinase isoforms. Notably, we
have discovered a new case of histone mark (PTM) crosstalk—an apparent gatekeeper function for histone H3
acetylation at Lys14 in blocking LSD1 demethylation of H3 methyl-Lys4. In the next phase of our research
program, we will develop and apply new chemical methods to more broadly understand the biology of histone
mark crosstalk using protein semisynthesis, gene editing, structural approaches, and “cut and paste” mass
spec proteomics. We will explore structural and functional features of enzymatic nucleosome interactions in the
context of LSD1, PRC1, SAGA, HDAC1, and Sirt6 complexes in part by incorporating chemical warheads into
designer nucleosomes. In addition, we will apply a newly engineered version of sortase to isolate histone H3
tails from cellular chromatin to quantitatively readout patterns of PTMs in different cell types and in response to
pharmacological agents by employing tandem mass tag mass spectrometry. Upon completion of this research
effort, our findings will broaden the knowledge of how histone Lys modifications are “written” and “erased” and
how specific PTM patterns regulate gene expression and cell fate. Moreover, these studies should pave the
way for new therapeutic strategies to combat epigenetic dysregulation in various diseases. This research
program will also enable the training of the next generation of biochemical investigators interested in protein
science and guided by a commitment to diversity and inclusion.

## Key facts

- **NIH application ID:** 10818361
- **Project number:** 5R35GM149229-02
- **Recipient organization:** BRIGHAM AND WOMEN'S HOSPITAL
- **Principal Investigator:** PHILIP A COLE
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $447,500
- **Award type:** 5
- **Project period:** 2023-04-01 → 2028-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10818361

## Citation

> US National Institutes of Health, RePORTER application 10818361, Chemical Approaches to Understanding Reversible Lysine Modifications (5R35GM149229-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10818361. Licensed CC0.

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