Sirtuin‐Derived Covalent Binder for the Selective Recognition of Protein Crotonylation

Ji, Yan-Li; Sun, Lin; Chen, Yao; Qin, Hongqiang; Xuan, Weimin

doi:10.1002/anie.202205522

Cited by 5 publications

(7 citation statements)

References 53 publications

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“…The recalcitrance of K Cr to react with thiols is reflected by the covalent capture of proteins containing it by phosphine-containing nucleophilic probes . However, an engineered sirtuin containing an unnatural thiol amino acid does covalently capture K Cr , Covalent modification of HDAC1 by K MP -modified histone proteins in cells could result in aberrant gene expression.…”

Section: Discussionmentioning

confidence: 99%

“…35 However, an engineered sirtuin containing an unnatural thiol amino acid does covalently capture K Cr . 36 N-Formyllysine (K Fo ), which is refractory to repair by HDAC1, also does not react with cysteine residues in proteins. 9,13 Covalent modification of HDAC1 by K MP -modified histone proteins in cells could result in aberrant gene expression.…”

Section: Hdac1 Competitive Inhibitor Prevents Covalent Modification B...mentioning

confidence: 99%

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Histone Deacetylase 1 Inhibition by Peptides Containing a DNA Damage-Induced, Nonenzymatic, Histone Covalent Modification

Jacinto

Greenberg

2023

Biochemistry

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Treatment of HeLa cells with the DNA damaging agent, bleomycin (BLM), results in the formation of a nonenzymatic 5-methylene-2-pyrrolone histone covalent modification on lysine residues (KMP). KMP is much more electrophilic than other N-acyllysine covalent modifications and post-translational modifications, including N-acetyllysine (KAc). Using histone peptides containing KMP, we show that this modification inhibits the class I histone deacetylase, HDAC1, by reacting with a conserved cysteine (C261) located near the active site. HDAC1 is inhibited by histone peptides whose corresponding N-acetylated sequences are known deacetylation substrates, but not one containing a scrambled sequence. The HDAC1 inhibitor, trichostatin A, competes with covalent modification by the KMP-containing peptides. HDAC1 is also covalently modified by a KMP-containing peptide in a complex milieu. These data indicate that peptides containing KMP are recognized by HDAC1 and are bound in the active site. The effects on HDAC1 indicate that KMP formation in cells may contribute to the biological effects of DNA damaging agents, such as BLM, that form this nonenzymatic covalent modification.

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Section: Discussionmentioning

confidence: 99%

Section: Hdac1 Competitive Inhibitor Prevents Covalent Modification B...mentioning

confidence: 99%

Histone Deacetylase 1 Inhibition by Peptides Containing a DNA Damage-Induced, Nonenzymatic, Histone Covalent Modification

Jacinto

Greenberg

2023

Biochemistry

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“…14,15 We are interested in designing sirtuin variants to address certain challenges in biological studies. Examples including sirtuin variants that could selectively cross-link with crotonylated lysine residues on proteins 16 and variants that exhibit orthogonal deacylase activities. 17 replacing an essential lysyl residue with N ε -acetyl-L-lysine (AcK) via GCE technology.…”

Section: ■ Introductionmentioning

confidence: 99%

A Sirtuin-Dependent T7 RNA Polymerase Variant

Wang,

Ji,

Sun

et al. 2023

ACS Synth. Biol.

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Transcriptional regulation is of great significance for cells to maintain homeostasis and, meanwhile, represents an innovative but less explored means to control biological processes in synthetic biology and bioengineering. Herein we devised a T7 RNA polymerase (T7RNAP) variant through replacing an essential lysine located in the catalytic core (K631) with N εacetyl-L-lysine (AcK) via genetic code expansion. This T7RNAP variant requires the deacetylase activity of NAD-dependent sirtuins to recover its enzymatic activities and thereby sustains sirtuindependent transcription of the gene of interest in live cells including bacteria and mammalian cells as well as in in vitro systems. This T7RNAP variant could link gene transcription to sirtuin expression and NAD availability, thus holding promise to support some relevant research.

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“…[8] And recently, Xuan et al developed a covalent binder for crotonylation by using a genetically thiol-coding bacterial sirtuin (CobB) to selectively recognize and react with Kcr sites in a unique NAD + dependent manner. [9] Here, we envisioned the unique α, β-unsaturated functionality of crotonyl as a potentially suitable site to enable Michael addition with cysteines of Kcr-interacting proteins in vicinity. However, the reactivity of Kcr was sterically hindered by the γ methyl group and not likely to occur.…”

Section: Introductionmentioning

confidence: 99%

“…And recently, Xuan et al . developed a covalent binder for crotonylation by using a genetically thiol‐coding bacterial sirtuin (CobB) to selectively recognize and react with Kcr sites in a unique NAD + dependent manner [9] …”

Section: Introductionmentioning

confidence: 99%

Development of Lysine Crotonyl‐Mimic Probe to Covalently Identify H3K27Cr Interacting Proteins

Guo,

Wang,

et al. 2023

Chemistry A European J

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Histone lysine crotonylation (Kcr) is one newly discovered acylation modification and regulates numerous pathophysiological processes. The binding affinity between Kcr and its interacting proteins is generally weak, which makes it difficult to effectively identify Kcr‐interacting partners. Changing the amide of crotonyl to an ester increased reactivity with proximal cysteines and retained specificity for Kcr antibody. We then designed the probe “H3g27Cr” by incorporating the ester functionality into a H3K27 peptide. Using this probe, we successfully identified multiple Kcr‐interacting partners including STAT3, which had not reported previously. Further experiments suggested that STAT3 possibly could form complexes with Histone deacetylase HDACs to downregulate the acetylation and crotonylation of Histone H3K27. Our unique design provided intriguing tools to further explore Kcr‐interacting proteins and elucidate their working mechanisms.

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Sirtuin‐Derived Covalent Binder for the Selective Recognition of Protein Crotonylation

Cited by 5 publications

References 53 publications

Histone Deacetylase 1 Inhibition by Peptides Containing a DNA Damage-Induced, Nonenzymatic, Histone Covalent Modification

Histone Deacetylase 1 Inhibition by Peptides Containing a DNA Damage-Induced, Nonenzymatic, Histone Covalent Modification

A Sirtuin-Dependent T7 RNA Polymerase Variant

Development of Lysine Crotonyl‐Mimic Probe to Covalently Identify H3K27Cr Interacting Proteins

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