Double quick, double click reversible peptide “stapling”

Abstract

A versatile, rapid and reversible approach to constrain peptides in a bioactive helical conformation and bearing a functional handle for inhibition of protein–protein interactions is described.

Bioconjugation with Maleimides: A Useful Tool for Chemical Biology

“…Although challenging, the use of dibromomaleimides hasa llowed the production of stable bioconjugates such as, structurally intact salmonc alcitonin with comb polymers derived from PEG methacrylic monomers, [51] or the ability to reversibly create constrained RNase Sp eptide sequence variants bearing cysteine( Cys) or homocysteine (hCys) aminoa cids. [52] Jackson and co-workersu sed N-6-caproamide-dibromomaleimides to prepareh omogeneous trastuzumab-MMAF and anti-CD98-MMAF constructs (68 and 69,r espectively,[ DAR] % 4), featuring improved in vivo efficacy when compared to constructs assem-Scheme12. Controlled functionalization of somatostatinusing aryloxymaleimides to yieldasingle modified conjugate (58)a nd one-pot sequential dual functionalization with aryloxymaleimidesa nd bromomaleimides to achieve dual modified (60)bioconjugates.…”

“…Eur.J. 2019,25,[43][44][45][46][47][48][49][50][51][52][53][54][55][56][57][58][59] www.chemeurj.org Given the reversible nature of 5MP-based bioconjugations, they have been showcased in protein pull-down strategies (Scheme 22 C). As olid support featuring primary amines in its surfacew as condensed with a5 MP precursor 101 to form immobilized 5MP 102.T hen, the first protein of interest (IlvN) was quantitatively immobilizedi nt he solids upport (< 10 min), while the remaining unreacted 5MP sites were capped with BME, ahead of incubation with ilvB (4 8C, 1h).…”

Tuning the Binding Affinity and Selectivity of Perfluoroaryl‐Stapled Peptides by Cysteine‐Editing

“…This topic has been recently reviewed by Fairlie . Examples of thiol cross‐linking include the use of dibromomaleimide, dichloroacetone, 1‐,4‐dichlorotetrazine, 1,2,4,5‐tetrabromodurene, α,α′‐dibromo‐ m ‐xylene, trans ‐1,4‐dibromo‐2‐butene and cis ‐1,4‐dichloro‐2‐butene and perfluoroaryl reagents . Although significant attention has focussed on the nature of the cross‐linking electrophile, comparatively little, if any, attention has focussed on the cysteine residues, with the single exception of introducing homocysteine .…”

“…Examples of thiol cross‐linking include the use of dibromomaleimide, dichloroacetone, 1‐,4‐dichlorotetrazine, 1,2,4,5‐tetrabromodurene, α,α′‐dibromo‐ m ‐xylene, trans ‐1,4‐dibromo‐2‐butene and cis ‐1,4‐dichloro‐2‐butene and perfluoroaryl reagents . Although significant attention has focussed on the nature of the cross‐linking electrophile, comparatively little, if any, attention has focussed on the cysteine residues, with the single exception of introducing homocysteine . The distance between the cysteine residues has been explored and optimised, albeit in non‐helical systems, yet the stereochemistry of cysteine has not been taken into account, in terms of the consequences on biological activity and the position of key amino acid side chain residues.…”

“…In light of these observations, we employed an in silico modelling approach (Figure ) to understand the structural and conformational consequences of cysteine replacement. The apparent α‐helicity of 7 was initially measured using circular dichroism but produced poor results (see Supporting Information) due to the absorbance of the fluoroaryl moiety at 222 nm, which was consistent with previous literature reports . Molecular dynamics simulations were performed starting from homology models for the free peptides 1 – 6 .…”