Secrets of a covalent interaction for biomaterials and biotechnology: SpyTag and SpyCatcher

Abstract

SpyTag is a short peptide that forms an isopeptide bond upon encountering its protein partner SpyCatcher. This covalent peptide interaction is a simple and powerful tool for bioconjugation and extending what protein architectures are accessible. Here we review the origin and mechanism of SpyTag/SpyCatcher, focusing on recent innovative applications. Ligation of targeting-antibody with antigen provided a simple route to vaccine generation. SpyRings, from head-to-tail cyclisation, gave major enhancements in enzyme resilience. Linking multiple SpyCatchers gave dendrimers for T-cell activation or Spy networks forming hydrogels for stem cell culture. Synthetic biology applications include integrating amyloid biomaterials with living bacteria, for irreversible derivatisation of biofilms with enzymes or nanoparticles. We also discuss further opportunities to apply and enhance SpyTag/SpyCatcher technology.

Evolving Accelerated Amidation by SpyTag/SpyCatcher to Analyze Membrane Dynamics

“…We have developed the use of spontaneous amide bond formation by peptide tags as a simple, specific, and genetically‐encoded route to lock protein units together 2. This technology, particularly the SpyTag/SpyCatcher pair, has been used in diverse applications including biomaterials, next‐generation sequencing, enzyme stabilization, and vaccine development 1a, 3. A key limitation has been relatively slow reaction at cellular expression levels.…”

Nouvelles stratégies d’innovations vaccinales et leurs application en médecine vétérinaire

“…Ces deux briques s'auto-assemblent spontanément pour former un pont isopeptidique de manière quasi-irréversible. La liaison est stable entre +4 et 37°C, dans une gamme de pH de 5 à 8, dans toutes sortes de milieux et même en présence de certains détergents (Li et al 2014;Reddington & Howarth, 2015). En recouvrant la surface de nanoparticules avec l'une des molécules de la paire SpyTag / SpyCatcher, et en attachant l'autre molécule de la paire sur les antigènes d'intérêt, il suffira de mélanger les deux pour créer un vaccin extrêmement stable, et ciblant directement les cellules dendritiques, qui sont programmées pour détecter de telles structures particulaires répétitives (figure 1).…”

Ethyl({[acryloyl(furan-2-ylmethyl)amino]acetyl}amino)acetate

“…They are the central framework of all biological events governing aspects of structure, transport, and catalytic pathways in nature [1]. Created intricately in a highly specific fashion, the functions of proteins and peptides in the living system have been a wonder and an inspiration in synthetic chemistry [2].…”