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

SNAP- 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 rev...

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Bibliographic Details
Published in:Journal of enzyme inhibition and medicinal chemistry 2021-01, Vol.36 (1), p.85-97
Main Authors: Merlo, Rosa, Caprioglio, Diego, Cillo, Michele, Valenti, Anna, Mattossovich, Rosanna, Morrone, Castrese, Massarotti, Alberto, Rossi, Franca, Miggiano, Riccardo, Leonardi, Antonio, Minassi, Alberto, Perugino, Giuseppe
Format: Article
Language:English
Subjects:
Azides - chemical synthesis
Azides - chemistry
biotechnology
Brief Report
click chemistry
DNA Modification Methylases - chemistry
DNA Modification Methylases - metabolism
enzymatic reaction
Fluorescent Dyes - chemical synthesis
Fluorescent Dyes - chemistry
HEK293 Cells
Humans
Models, Molecular
Molecular Structure
protein labelling
protein-tag
Short Communication
Structure-Activity Relationship
Substrate Specificity
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Summary:SNAP- 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 , by using a BG-substrate containing an azide group appropriately distanced by a spacer from the benzyl ring. The SNAP- and its relative thermostable version ( OGT- ) proved to be very active on this substrate. The stability of these 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 .
ISSN:1475-6366
1475-6374
1475-6374
DOI:10.1080/14756366.2020.1841182