Co‐translational Installation of Posttranslational Modifications by Non‐canonical Amino Acid Mutagenesis

Niu, Wei; Guo, Jiantao

doi:10.1002/cbic.202300039

Cited by 7 publications

(4 citation statements)

References 195 publications

(334 reference statements)

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“…172 The applications of these AcK-OTSs in determining the roles of lysine acetylation in regulating specific proteins, from histones to nonhistone proteins, have been nicely reviewed recently. 36,38,40,173 There is one potential problem when using AcK-OTSs for in vivo studies. Due to existence of deacetylases, site-specifically incorporated acetyllysine residues by GCE undergo deacetylation, especially in yeast or mammalian cells, and the stoichiometry of acetylation at target sites varies from batch to batch to make inconsistence among replicates.…”

Section: Lysine Acetylationmentioning

confidence: 99%

“…Furthermore, because of the orthogonality of PylRS-derived OTSs in both bacteria and eukaryotes, genetic encoding of acetyllysine into proteins has been established in different organisms, including E. coli , , Salmonella , yeast, , mammalian cells, , stem cells, and animals . The applications of these AcK-OTSs in determining the roles of lysine acetylation in regulating specific proteins, from histones to nonhistone proteins, have been nicely reviewed recently. ,,, …”

Section: Lysine Modificationsmentioning

confidence: 99%

“…To fill in this gap, the genetic code expansion (GCE) strategy has been applied for PTM studies. − It introduces an orthogonal translation system (OTS) into living cells to genetically encode a noncanonical amino acid (ncAA) at a chosen site of target proteins. − The OTS is mainly composed of an engineered aminoacyl-tRNA synthetase (AARS), which recognizes the ncAA, and an engineered tRNA, which bears a specific anticodon to read an assigned codon in mRNA (Figure ). − The orthogonality of an OTS includes the following: First, the target ncAA cannot be recognized by host AARSs; Second, the engineered AARS cannot recognize canonical amino acids and cannot aminoacylate host tRNAs; Third, the engineered tRNA cannot be aminoacylated by host AARSs.…”

Section: Introductionmentioning

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%

Section: Introductionmentioning

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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“…In many cases, these covalent modifications cannot easily be recapitulated in recombinant proteins, and as a result their functional significance can be difficult to elucidate . GCE offers a powerful approach to probe the role of PTMs, through selective introduction of ncAAs that are structural mimics of post-translationally modified residues. − …”

Section: Probing Enzyme Mechanisms With Noncanonical Amino Acidsmentioning

confidence: 99%

Noncanonical Amino Acids in Biocatalysis

Birch-Price,

Hardy,

Lister

et al. 2024

Chem. Rev.

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In recent years, powerful genetic code reprogramming methods have emerged that allow new functional components to be embedded into proteins as noncanonical amino acid (ncAA) side chains. In this review, we will illustrate how the availability of an expanded set of amino acid building blocks has opened a wealth of new opportunities in enzymology and biocatalysis research. Genetic code reprogramming has provided new insights into enzyme mechanisms by allowing introduction of new spectroscopic probes and the targeted replacement of individual atoms or functional groups. NcAAs have also been used to develop engineered biocatalysts with improved activity, selectivity, and stability, as well as enzymes with artificial regulatory elements that are responsive to external stimuli. Perhaps most ambitiously, the combination of genetic code reprogramming and laboratory evolution has given rise to new classes of enzymes that use ncAAs as key catalytic elements. With the framework for developing ncAA-containing biocatalysts now firmly established, we are optimistic that genetic code reprogramming will become a progressively more powerful tool in the armory of enzyme designers and engineers in the coming years.

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Phosphorylation of cytochrome c at tyrosine 48 finely regulates its binding to the histone chaperone SET/TAF‐Iβ in the nucleus

Tamargo‐Azpilicueta,

Casado‐Combreras,

Giner‐Arroyo

et al. 2024

Protein Science

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Post‐translational modifications (PTMs) of proteins are ubiquitous processes present in all life kingdoms, involved in the regulation of protein stability, subcellular location and activity. In this context, cytochrome c (Cc) is an excellent case study to analyze the structural and functional changes induced by PTMS as Cc is a small, moonlighting protein playing different roles in different cell compartments at different cell‐cycle stages. Cc is actually a key component of the mitochondrial electron transport chain (ETC) under homeostatic conditions but is translocated to the cytoplasm and even the nucleus under apoptotic conditions and/or DNA damage. Phosphorylation does specifically alter the Cc redox activity in the mitochondria and the Cc non‐redox interaction with apoptosis‐related targets in the cytoplasm. However, little is known on how phosphorylation alters the interaction of Cc with histone chaperones in the nucleus. Here, we report the effect of Cc Tyr48 phosphorylation by examining the protein interaction with SET/TAF‐Iβ in the nuclear compartment using a combination of molecular dynamics simulations, biophysical and structural approaches such as isothermal titration calorimetry (ITC) and nuclear magnetic resonance (NMR) and in cell proximity ligation assays. From these experiments, we infer that Tyr48 phosphorylation allows a fine‐tuning of the Cc‐mediated inhibition of SET/TAF‐Iβ histone chaperone activity in vitro. Our findings likewise reveal that phosphorylation impacts the nuclear, stress‐responsive functions of Cc, and provide an experimental framework to explore novel aspects of Cc post‐translational regulation in the nucleus.

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Co‐translational Installation of Posttranslational Modifications by Non‐canonical Amino Acid Mutagenesis

Cited by 7 publications

References 195 publications

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

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

Noncanonical Amino Acids in Biocatalysis

Phosphorylation of cytochrome c at tyrosine 48 finely regulates its binding to the histone chaperone SET/TAF‐Iβ in the nucleus

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