Fluorescent Amino Acid Initiated <i>de novo</i> Cyclic Peptides for the Label‐Free Assessment of Cell Permeability**

Wu, Yinghai; Bertran, M. Teresa; Rowley, James; Calder, Ewen D. D.; Joshi, Dhira; Walport, Louise J.

doi:10.1002/cmdc.202100315

Cited by 6 publications

(2 citation statements)

References 22 publications

(38 reference statements)

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“…Although polypeptide translation typically begins with f Met in nature, the initiation event tolerates a wide variety of npMs, even in the absence of tRNA fMet CAU engineering. ,, Examples translated in the FIT and RaPID systems include l -npMs, − d -AAs, , N α -acyl-AAs, , N α -alkyl-AAs, , γ 4 -AAs, δ 5 -AAs, ε 6 -AAs, ζ 7 -AAs, farnesyl-AAs, malonyl-AAs, biotinylated AAs, ,− (amino)benzoic acids, ,,,,− heteroaromatics, oligopeptides, , foldamers, − bipyridines, and carborane . As mentioned in section , the engineered tRNA iniP has been used to initiate translation with relatively more unreactive substrates such as a β 3 -AA and a cyclic γ 2,4 -AA .…”

Section: Applications Of Polypeptides Containing Ribosomally Translat...mentioning

confidence: 99%

Engineering tRNAs for the Ribosomal Translation of Non-proteinogenic Monomers

Sigal,

Matsumoto,

Beattie

et al. 2024

Chem. Rev.

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Ribosome-dependent protein biosynthesis is an essential cellular process mediated by transfer RNAs (tRNAs). Generally, ribosomally synthesized proteins are limited to the 22 proteinogenic amino acids (pAAs: 20 L-α-amino acids present in the standard genetic code, selenocysteine, and pyrrolysine). However, engineering tRNAs for the ribosomal incorporation of non-proteinogenic monomers (npMs) as building blocks has led to the creation of unique polypeptides with broad applications in cellular biology, material science, spectroscopy, and pharmaceuticals. Ribosomal polymerization of these engineered polypeptides presents a variety of challenges for biochemists, as translation efficiency and fidelity is often insufficient when employing npMs. In this Review, we will focus on the methodologies for engineering tRNAs to overcome these issues and explore recent advances both in vitro and in vivo. These efforts include increasing orthogonality, recruiting essential translation factors, and creation of expanded genetic codes. After our review on the biochemical optimizations of tRNAs, we provide examples of their use in genetic code manipulation, with a focus on the in vitro discovery of bioactive macrocyclic peptides containing npMs. Finally, an analysis of the current state of tRNA engineering is presented, along with existing challenges and future perspectives for the field.

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Section: Applications Of Polypeptides Containing Ribosomally Translat...mentioning

confidence: 99%

Engineering tRNAs for the Ribosomal Translation of Non-proteinogenic Monomers

Sigal,

Matsumoto,

Beattie

et al. 2024

Chem. Rev.

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show abstract

“…In that line, cyclopeptides are promising candidates for efficient anion sensors and anion transporters in vivo due to the abundance and orientation of anion-binding groups, including amide protons, and resistance to biodegradation when compared to their linear analogues. One of the most sensitive methods used for the detection of anion-receptor complex formation in solution is fluorimetry [15,[18][19][20][21][22]. The method is fast, reliable, and gives a multivariate response that is well suited for stability constants determination.…”

Section: Introductionmentioning

confidence: 99%

Anion-Sensing Properties of Cyclopentaphenylalanine

et al. 2022

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Cyclic pentaphenylalanine was studied as an efficient anion sensor for halides, thiocyanate and oxoanions in acetonitrile and methanol. Stability constants of the corresponding complexes were determined by means of fluorimetric, spectrophotometric, 1H NMR, and microcalorimetric titrations. A detailed structural overview of receptor–anion complexes was obtained by classical molecular dynamics (MD) simulations. The results of 1H NMR and MD studies indicated that the bound anions were coordinated by the amide groups of cyclopeptide, as expected. Circular dichroism (CD) titrations were also carried out in acetonitrile. To the best of our knowledge, this is the first example of the detection of anion binding by cyclopeptide using CD spectroscopy. The CD spectra were calculated from the structures obtained by MD simulations and were qualitatively in agreement with the experimental data. The stoichiometry of almost all complexes was 1:1 (receptor:anion), except for dihydrogen phosphate where the binding of dihydrogen phosphate dimer was observed in acetonitrile. The affinity of the cyclopeptide receptor was correlated with the structure of anion coordination sphere, as well as with the solvation properties of the examined solvents.

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Fluorescent Amino Acid Initiated de novo Cyclic Peptides for the Label‐Free Assessment of Cell Permeability**

Bertran

Rowley

et al. 2021

ChemMedChem

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The major obstacle in applying peptides to intracellular targets is their low inherent cell permeability. Standard approaches to attach a fluorophore (e. g. FITC, TAMRA) can change the physicochemical properties of the parent peptide and influence their ability to penetrate and localize in cells. We report a label‐free strategy for evaluating the cell permeability of cyclic peptide leads. Fluorescent tryptophan analogues 4‐cyanotryptophan (4CNW) and β‐(1‐azulenyl)‐L‐alanine (AzAla) were incorporated into in vitro translated macrocyclic peptides by initiator reprogramming. We then demonstrate these efficient blue fluorescent emitters are good tools for monitoring peptide penetration into cells.

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Fluorescent Amino Acid Initiated de novo Cyclic Peptides for the Label‐Free Assessment of Cell Permeability**

Cited by 6 publications

References 22 publications

Engineering tRNAs for the Ribosomal Translation of Non-proteinogenic Monomers

Engineering tRNAs for the Ribosomal Translation of Non-proteinogenic Monomers

Anion-Sensing Properties of Cyclopentaphenylalanine

Fluorescent Amino Acid Initiated de novo Cyclic Peptides for the Label‐Free Assessment of Cell Permeability**

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