Scalable synthesis and coupling of quaternary α-arylated amino acids: α-aryl substituents are tolerated in α-helical peptides

Leonard, Daniel J.; Zieleniewski, Francis; Wellhöfer, Isabelle; Baker, Emily G.; Ward, John W.; Woolfson, Derek N.; Clayden, Jonathan

doi:10.1039/d1sc01378e

Cited by 5 publications

(6 citation statements)

References 27 publications

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“…For instance, we replaced the remaining large hydrophobic Ile residues with Ala in a 3-heptad HERD background, to resemble a known monomeric α helix (HERD-Ctrl1–GFP and Ctrl2, Fig. 2a,b ) 51 . Also, to eliminate the amphipathicity of the HRs, we scrambled their sequences (HERD-Ctrl3–GFP).…”

Section: Resultsmentioning

confidence: 99%

Assembling membraneless organelles from de novo designed proteins

Hilditch,

Romanyuk,

Cross

et al. 2023

Nat. Chem.

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Recent advances in de novo protein design have delivered a diversity of discrete de novo protein structures and complexes. A new challenge for the field is to use these designs directly in cells to intervene in biological processes and augment natural systems. The bottom-up design of self-assembled objects such as microcompartments and membraneless organelles is one such challenge. Here we describe the design of genetically encoded polypeptides that form membraneless organelles in Escherichia coli. To do this, we combine de novo α-helical sequences, intrinsically disordered linkers and client proteins in single-polypeptide constructs. We tailor the properties of the helical regions to shift protein assembly from arrested assemblies to dynamic condensates. The designs are characterized in cells and in vitro using biophysical methods and soft-matter physics. Finally, we use the designed polypeptide to co-compartmentalize a functional enzyme pair in E. coli, improving product formation close to the theoretical limit.

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Section: Resultsmentioning

confidence: 99%

Assembling membraneless organelles from de novo designed proteins

Hilditch,

Romanyuk,

Cross

et al. 2023

Nat. Chem.

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“…To further utilize the afforded α,α-disubstituted α-AAs in peptide synthesis especially incorporating this unit into the peptides via Fmoc solid-phase peptide synthesis (SPPS), a more practical and scalable approach to synthesize α,α-disubstituted α-AAs bearing aryl groups was introduced by the same group. The stability and tolerance of α,α-disubstituted α-AAs in a α-helical peptide was verified, which showcased the potential utility of such α,α-disubstituted α-AAs via arylation of other proteinogenic α-AAs 37 . Beside the arylation of the general amino acids, the same group also disclosed the α-alkenylation of general α-amino acids using similar strategy to extend the applications of this approach 38 .…”

Section: Synthesis Of αα-Disubstituted Amino Acids Via the C-c Bond F...mentioning

confidence: 85%

“…Another conceptual improvement is to extend the coupling partners towards general alkyl halides 12 and versatile tertiary hydrocarbons which were highly challenging in previous reports 36 .The key to success is the radical-radical coupling of two radical species that are obtained through a one-electron process. α-Functionalization of general α-AAs was also reported recently to realize various α,α-disubstituted α-AAs, especially extending to the arylation and alkenylation of α-amino acids via multi-step pathways [37][38][39] . The key step of the C-C bond formation is the rearrangement reaction to realize the arylation of the amino acid α-center in a diastereoselective manner.…”

Section: Conceptual Overview For the Synthesis Of αα-Disubstituted α-Aasmentioning

confidence: 99%

Challenges and recent advancements in the synthesis of α,α-disubstituted α-amino acids

Zhang,

Vanderghinste,

Wang

et al. 2024

Nat Commun

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Abstractα,α-Disubstituted α-amino acids (α-AAs) have improved properties compared to other types of amino acids. They serve as modifiers of peptide conformation and as precursors of bioactive compounds. Therefore, it has been a long-standing goal to construct this highly valuable scaffold efficiently in organic synthesis and drug discovery. However, access to α,α-disubstituted α-AAs is highly challenging and largely unexplored due to their steric constraints. To overcome these, remarkable advances have been made in the last decades. Emerging strategies such as synergistic enantioselective catalysis, visible-light-mediated photocatalysis, metal-free methodologies and CO2 fixation offer new avenues to access the challenging synthesis of α,α-disubstituted α-AAs and continuously bring additional contributions to this field. This review article aims to provide an overview of the recent advancements since 2015 and discuss existing challenges for the synthesis of α,α-disubstituted α-AAs and their derivatives.

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“…For instance, we replaced the remaining large hydrophobic Ile residues with Ala in a 3 heptad HERD background, to resemble a known monomeric α helix (HERD-Ctrl1-GFP & Ctrl2). 48 To eliminate the amphipathicity of the HRs, we scrambled their sequences (HERD-Ctrl3-GFP). And, we introduced helix-breaking mutations into the HRs, e.g.…”

Section: Nascent Helicity Helps Drive the Condensation Of Herd-22-gfpmentioning

confidence: 99%

Assembling membraneless organelles fromde novodesigned proteins

Hilditch

Romanyuk

Cross

et al. 2023

Preprint

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Recent advances inde novoprotein design have delivered a diversity of discretede novoprotein structures and complexes. A new challenge for the field is to use these designs directly in cells to intervene in biological process and augment natural systems. The bottom-up design of self-assembled objects like microcompartments and membraneless organelles is one such challenge, which also presents opportunities for chemical and synthetic biology. Here, we describe the design of genetically encoded polypeptides that form membraneless organelles inEscherichia coli(E. coli). To do this, we combinede novoα-helical sequences, intrinsically disordered linkers, and client proteins in single-polypeptide constructs. We tailor the properties of the helical regions to shift protein assembly from diffusion-limited assemblies to dynamic condensates. The designs are characterised in cells andin vitrousing biophysical and soft-matter physics methods. Finally, we use the designed polypeptide to co-compartmentalise a functional enzyme pair inE. coli.

show abstract

Scalable synthesis and coupling of quaternary α-arylated amino acids: α-aryl substituents are tolerated in α-helical peptides

Abstract: Quaternary amino acids are important tools for the modification and stabilisation of peptide secondary structures. Here we describe a practical and scalable synthesis applicable to quaternary alpha-arylated amino acids (Q4As),...

Cited by 5 publications

References 27 publications

Assembling membraneless organelles from de novo designed proteins

Assembling membraneless organelles from de novo designed proteins

Challenges and recent advancements in the synthesis of α,α-disubstituted α-amino acids

Assembling membraneless organelles fromde novodesigned proteins

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