Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs.

ABSTRACT: Formylglycine-generating enzymes specifically oxidize cysteine within the consensus sequence CxPxR to C^α -formylglycine (FGly). This noncanonical electrophilic amino acid can subsequently be addressed selectively by bioorthogonal hydrazino-iso-Pictet-Spengler (HIPS) or Knoevenagel ligation to attach payloads like fluorophores or drugs to proteins to obtain a defined payload-to-protein ratio. However, the disadvantages of these conjugation techniques include the need for a large excess of conjugation building block, comparably low reaction rates and limited stability of FGly-containing proteins. Therefore, functionalized clickable HIPS and tandem Knoevenagel building blocks were synthesized, conjugated to small proteins (DARPins) and subsequently linked to strained alkyne-contain

SUBMITTER: Janson N 

PROVIDER: S-EPMC7756428 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature