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PreprintCited by 2Click Chemistry and ApplicationsRNA and protein synthesis mechanismsChemical Synthesis and Analysis

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

A Modular Genetic Code Expansion Approach to Site-Specific Lysine Acylations

Open access
Knecht, Pascal, Nguyen, Tuan-Anh, Lang, Kathrin·2025·Chem
DOI 10.1101/2025.10.03.680225

Preprint — published as 10.1016/j.chempr.2025.102919 · Paper page

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Abstract

Lysine acylations are a diverse class of post-translational modifications (PTMs) that dynamically regulate protein function. Access to homogenous, site-specifically modified proteins is essential for dissecting their molecular roles, yet remains challenging with traditional methods. Here we present a modular strategy that combines genetic code expansion (GCE) with a chemoselective amide bond-forming reaction to install a wide range of lysine acylations directly onto folded proteins. Key to this approach is the genetic encoding of N ε -methoxylysine, which reacts efficiently with N -methyliminodiacetyl (MIDA) acylboronates to generate diverse acyl lysine modifications from a single protein precursor, including several acyl PTMs not previously accessible through GCE. This mild and broadly applicable methodology enables systematic studies of lysine acylation on multiple target proteins. We demonstrate its utility by probing the effects of defined acyl modifications on enzymatic activity, protein-RNA interactions and deacylase-mediated regulation. Together, this platform establishes a versatile strategy to interrogate the functional consequences of this wide-spread class of PTMs.

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Metadata

Authors
Knecht, P., Nguyen, T.-A., & Lang, K.
Affiliations
ETH Zurich (CH)
Corresponding author
Lang, Kathrin
ORCID
Venue
Chem
Publisher
openRxiv
Published
2025-10-03
Volume
12
Issue
5
Pages
102919-102919
ISSN
2451-9294 (ISSN-L)
DOI
10.1101/2025.10.03.680225
OpenAlex
W7133318153
Cited by
2
References
1
Citations by year
2026: 2
FWCI
3.48
Type
journal article
Language
en
Open access
Licence
Locations
  • Chem (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • Repository for Publications and Research Data (ETH Zurich) (publishedVersion, cc-by)
  • ETH Zürich Research Collection (cc-by)
Funders
  • European Commission (HORIZON2020, 101003289)
  • Eidgenössische Technische Hochschule Zürich
  • Horizon 2020 Framework Programme
  • European Research Council
  • H2020 European Research Council (101003289)
Topics
  • Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Physical Sciences › Chemistry › Organic Chemistry › Chemical Synthesis and Analysis
Keywords
lysine acylation, genetic code expansion, site-specific protein modification, post-translational modification, RNA-protein interactions, enzyme activity

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