The SNAP-tag technology revised: an effective chemo-enzymatic approach by using a universal azide-based substrate

• Main Authors: Rosa Merlo (Author), Diego Caprioglio (Author), Michele Cillo (Author), Anna Valenti (Author), Rosanna Mattossovich (Author), Castrese Morrone (Author), Alberto Massarotti (Author), Franca Rossi (Author), Riccardo Miggiano (Author), Antonio Leonardi (Author), Alberto Minassi (Author), Giuseppe Perugino (Author)
• Format: Book
• Published: Taylor & Francis Group, 2021-01-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

SNAP-tag® is a powerful technology for the labelling of protein/enzymes by using benzyl-guanine (BG) derivatives as substrates. Although commercially available or ad hoc produced, their synthesis and purification are necessary, increasing time and costs. To address this limitation, here we suggest a revision of this methodology, by performing a chemo-enzymatic approach, by using a BG-substrate containing an azide group appropriately distanced by a spacer from the benzyl ring. The SNAP-tag® and its relative thermostable version (SsOGT-H5) proved to be very active on this substrate. The stability of these tags upon enzymatic reaction makes possible the exposition to the solvent of the azide-moiety linked to the catalytic cysteine, compatible for the subsequent conjugation with DBCO-derivatives by azide-alkyne Huisgen cycloaddition. Our studies propose a strengthening and an improvement in terms of biotechnological applications for this self-labelling protein-tag.

Item Description

1475-6366
1475-6374
10.1080/14756366.2020.1841182