Catalytic Enantioselective Smiles Rearrangement Enabled by the Directed Evolution of P450 Radical Aryl Migratases

Journal of the American Chemical Society, Год журнала: 2025, Номер unknown

Опубликована: Апрель 1, 2025

Despite its synthetic potential, catalytic enantioselective Smiles rearrangement has remained elusive. Through the directed evolution of P450 radical aryl migratases (P450Smiles's), we describe first example rearrangement. A range racemic N-arylsulfonyl-α-chloroamides could be transformed by P450Smiles in an enantioconvergent manner, affording acyclic amide products possessing all-carbon quaternary stereocenter with excellent chemo- and enantioselectivity. Both electron-rich electron-deficient substituents were compatible migrating group, demonstrating this P450-catalyzed is insensitive to electronic properties group. Importantly, our evolved variants capable overriding innate cyclization activity N-alkyl amidyl intermediate, allowing chemoselective reductive formation products. Classical molecular dynamics (MD) simulations revealed unusual enzyme-controlled chemoselectivity stems from restricted conformation within enzyme active site, disfavoring pathway. This new-to-nature biocatalytic asymmetric showcases potential enzymatic enantioselectivity control over highly reactive intermediates eluding small-molecule catalysts.

Язык: Английский

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Mechanistic Insights into GTP Hydrolysis by the RhoA Protein: Catalytic Impact of Glutamine Tautomerism

ACS Catalysis, Год журнала: 2025, Номер unknown, С. 4415 - 4428

Опубликована: Фев. 27, 2025

We present a systematic evaluation of different possible reaction mechanisms for GTP hydrolysis in RhoA, member the Ras superfamily enzymes that uses this to switch from an active inactive conformation. These are activated by presence GTPase activating protein (or GAP) forms intimate complex with residues two proteins site. have explored multidimensional reactional free energy landscape site formed RhoA and p50RhoGAP. Our molecular dynamics simulations show enzyme p50RhoGAP establishes catalytically important interactions phosphate groups through its so-called arginine finger (Arg85) also residue Gln63. This is key because it not only interacts nucleophilic water molecule but participates actively mechanism. Adaptive string method using hybrid quantum mechanics/molecular mechanics (QM/MM) potentials both tight-binding density functional Hamiltonians proceeds formation metaphosphate metastable species. Mechanistic proposals differ proton transfer rearrangements required form inorganic ion. discard solvent-assisted mechanism point participation Gln63 process means side chain tautomerism amide imide form. The recover requires phosphate, rate-limiting step process, barrier 20.2 kcal mol-1 at B3LYPD3/MM level, good agreement experimentally derived value. amide-imide could be relevant other enzymes, facilitating events mechanisms.

Язык: Английский

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Emergent Mechanisms in Biocatalysis

ACS Central Science, Год журнала: 2025, Номер unknown

Опубликована: Июнь 5, 2025

Язык: Английский

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