Synthesis, Biochemical Characterization, and Genetic Encoding of a 1,2,4‐Triazole Amino Acid as an Acetyllysine Mimic for Bromodomains of the BET Family

Kirchgäßner, Sören; Braun, Michael B.; Bartlick, Natascha; Koç, Cengiz; Reinkemeier, Christopher D.; Lemke, Edward A.; Stehle, Thilo; Schwarzer, Dirk

doi:10.1002/anie.202215460

Cited by 3 publications

(3 citation statements)

References 61 publications

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“…Recently, another stable mimic of acetyllysine, 1,2,4-triazole amino acid (ApmTri), has been site-specifically incorporated into proteins by WT MmPylRS in mammalian cells. 178 The ApmTri-containing proteins were shown to have similar binding affinity with other proteins to that of acetyllysinecontaining proteins. All these above-mentioned acetyllysine mimics offer alternative approaches when acetyllysine residues undergo rapid deacetylation in living cells.…”

Section: Lysine Acetylationmentioning

confidence: 99%

“…In this study, TFAcK-containing proteins were also demonstrated to be resistant to sirtuin deacetylases. Recently, another stable mimic of acetyllysine, 1,2,4-triazole amino acid (ApmTri), has been site-specifically incorporated into proteins by WT Mm PylRS in mammalian cells . The ApmTri-containing proteins were shown to have similar binding affinity with other proteins to that of acetyllysine-containing proteins.…”

Section: Lysine Modificationsmentioning

confidence: 99%

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Orthogonal Translation for Site-Specific Installation of Post-translational Modifications

Gan,

Fan

2024

Chem. Rev.

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Post-translational modifications (PTMs) endow proteins with new properties to respond to environmental changes or growth needs. With the development of advanced proteomics techniques, hundreds of distinct types of PTMs have been observed in a wide range of proteins from bacteria, archaea, and eukarya. To identify the roles of these PTMs, scientists have applied various approaches. However, high dynamics, low stoichiometry, and crosstalk between PTMs make it almost impossible to obtain homogeneously modified proteins for characterization of the site-specific effect of individual PTM on target proteins. To solve this problem, the genetic code expansion (GCE) strategy has been introduced into the field of PTM studies. Instead of modifying proteins after translation, GCE incorporates modified amino acids into proteins during translation, thus generating site-specifically modified proteins at target positions. In this review, we summarize the development of GCE systems for orthogonal translation for site-specific installation of PTMs.

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Section: Lysine Acetylationmentioning

confidence: 99%

Section: Lysine Modificationsmentioning

confidence: 99%

Orthogonal Translation for Site-Specific Installation of Post-translational Modifications

Gan,

Fan

2024

Chem. Rev.

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“…In addition to the natural modification, lysine acetylation mimics, such as thioacetylation, trifluoroacetylation, and 1,2,4‐triazole, have also be designed and integrated into proteins in the forms of ε‐ N ‐thioacetyl‐lysine (TAcK; Venkat et al, 2017), ε‐ N ‐trifluoroacetyl‐lysine (TFAcK; Zhang et al, 2018), and 1,2,4‐triazole amino acid (ApmTri; Kirchgäßner et al, n.d.), respectively (Figure 2a). All of these modifications are non‐hydrolyzable and resistant to deacetylases, providing alternative approaches to access stable and homogenous proteins bearing acetylation mimics.…”

Section: Lysine Modificationsmentioning

confidence: 99%

Functional analysis of protein post‐translational modifications using genetic codon expansion

et al. 2023

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Post-translational modifications (PTMs) of proteins not only exponentially increase the diversity of proteoforms, but also contribute to dynamically modulating the localization, stability, activity, and interaction of proteins. Understanding the biological consequences and functions of specific PTMs has been challenging for many reasons, including the dynamic nature of many PTMs and the technical limitations to access homogenously modified proteins. The genetic code expansion technology has emerged to provide unique approaches for studying PTMs. Through site-specific incorporation of unnatural amino acids (UAAs) bearing PTMs or their mimics into proteins, genetic code expansion allows the generation of homogenous proteins with site-specific modifications and atomic resolution both in vitro and in vivo. With this technology, various PTMs and mimics have been precisely introduced into proteins. In this review, we summarize the UAAs and approaches that have been recently developed to site-specifically install PTMs and their mimics into proteins for functional studies of PTMs.

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Role of the histone variant H2A.Z.1 in memory, transcription, and alternative splicing is mediated by lysine modification

Reda,

Hategan,

McLean

et al. 2024

Neuropsychopharmacol.

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Creating long-lasting memories requires learning-induced changes in gene expression, which are impacted by epigenetic modifications of DNA and associated histone proteins. Post-translational modifications (PTMs) of histones are key regulators of transcription, with different PTMs producing unique effects on gene activity and behavior. Although recent studies implicate histone variants as novel regulators of memory, effects of PTMs on the function of histone variants are rarely considered. We previously showed that the histone variant H2A.Z suppresses memory, but it is unclear if this role is impacted by H2A.Z acetylation, a PTM that is typically associated with positive effects on transcription and memory. To answer this question, we used a mutation approach to manipulate acetylation on H2A.Z without impacting acetylation of other histone types. Specifically, we used adeno-associated virus (AAV) constructs to overexpress mutated H2A.Z.1 isoforms that either mimic acetylation (acetyl-mimic) by replacing lysines 4, 7 and 11 with glutamine (KQ), or H2A.Z.1 with impaired acetylation (acetyl-defective) by replacing the same lysines with alanine (KA). Expressing the H2A.Z.1 acetyl-mimic (H2A.Z.1KQ) improved memory under weak learning conditions, whereas expressing the acetyl-defective H2A.Z.1KA generally impaired memory, indicating that the effect of H2A.Z.1 on memory depends on its acetylation status. RNA sequencing showed that H2A.Z.1KQ and H2A.Z.1KA uniquely impact the expression of different classes of genes in both females and males. Specifically, H2A.Z.1KA preferentially impacts genes involved in synaptic function, suggesting that acetyl-defective H2A.Z.1 impairs memory by altering synaptic regulation. Finally, we describe, for the first time, that H2A.Z is also involved in alternative splicing of neuronal genes, whereby H2A.Z depletion, as well as expression of H2A.Z.1 lysine mutants influence transcription and splicing of different gene targets, suggesting that H2A.Z.1 can impact behavior through effects on both splicing and gene expression. This is the first study to demonstrate that direct manipulation of H2A.Z post-translational modifications regulates memory, whereby acetylation adds another regulatory layer by which histone variants can fine tune higher brain functions through effects on gene expression and splicing.

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Synthesis, Biochemical Characterization, and Genetic Encoding of a 1,2,4‐Triazole Amino Acid as an Acetyllysine Mimic for Bromodomains of the BET Family

Cited by 3 publications

References 61 publications

Orthogonal Translation for Site-Specific Installation of Post-translational Modifications

Orthogonal Translation for Site-Specific Installation of Post-translational Modifications

Functional analysis of protein post‐translational modifications using genetic codon expansion

Role of the histone variant H2A.Z.1 in memory, transcription, and alternative splicing is mediated by lysine modification

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