# Develop General Methods for the Synthesis of Proteins with Posttranslational Lysine Modifications

> **NIH NIH R01** · TEXAS A&M UNIVERSITY · 2021 · $350,805

## Abstract

PROJECT SUMMARY/ABSTRACT
 Among all 20 native amino acids in proteins, lysine (Lys) undergoes the most diverse forms of
posttranslational modifications (PTMs). The unique nucleophilicity of its side chain amine allows Lys to be
selectively modified with several types of alkylation and a number of small-molecule and protein acylations.
These PTMs, especially in eukaryotes, regulate enzyme activities, interactions of proteins with their partners,
cellular localization of proteins, and protein metabolism. Abnormality of these PTMs correlates with the
development of many diseases. Although important, biochemical studies of Lys PTMs are cumbersome due to
the difficulty to synthesize proteins with them. Several methods have been developed for the synthesis of
proteins with Lys PTMs. However, they cannot be generally applied. With an overall objective to formulate
straightforward methods that can be generally applied for the synthesis of proteins with Lys PTMs, the current
application will focus on the development of amber suppression-based noncanonical amino acid (ncAA)
mutagenesis methods in combination with chemical transformation for the installation of Lys PTMs into
proteins. Three specific aims will be pursued: 1) Develop enhanced amber suppression-based ncAA
mutagenesis methods for the recombinant synthesis of proteins with Lys alkylations and small-molecule
acylations; 2) Develop amber suppression-based ncAA mutagenesis methods for the synthesis of proteins with
ubiquitin and ubiquitin-like protein modifications; and 3) Formulate recombinant strategies in conjunction with
biocompatible reactions to synthesize proteins installed site-specifically with two different Lys PTMs. The
successful completion of the proposed study will make available straightforward methods for the synthesis of
proteins with most Lys PTMs for their functional investigation.

## Key facts

- **NIH application ID:** 10063529
- **Project number:** 5R01GM127575-03
- **Recipient organization:** TEXAS A&M UNIVERSITY
- **Principal Investigator:** Wenshe Ray Liu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $350,805
- **Award type:** 5
- **Project period:** 2019-01-15 → 2022-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10063529

## Citation

> US National Institutes of Health, RePORTER application 10063529, Develop General Methods for the Synthesis of Proteins with Posttranslational Lysine Modifications (5R01GM127575-03). Retrieved via AI Analytics 2026-06-11 from https://api.ai-analytics.org/grant/nih/10063529. Licensed CC0.

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