Insider information on successful covalent protein coupling with help from SpyBank

New biological properties can stem from the freedom to link, multimerize, or multiplex protein building blocks. The peptide SpyTag on one protein irreversibly reacts with SpyCatcher on another protein, through spontaneous isopeptide bond formation. Reaction is specific in a wide range of cellular en...

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Bibliographic Details
Main Authors: Keeble, A, Howarth, M
Format: Book section
Language:English
Published: Elsevier 2019
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author Keeble, A
Howarth, M
author_facet Keeble, A
Howarth, M
author_sort Keeble, A
collection OXFORD
description New biological properties can stem from the freedom to link, multimerize, or multiplex protein building blocks. The peptide SpyTag on one protein irreversibly reacts with SpyCatcher on another protein, through spontaneous isopeptide bond formation. Reaction is specific in a wide range of cellular environments and all components are genetically encoded, making this chemistry accessible to molecular biologists. SpyTag/SpyCatcher has been widely used for enzyme immobilization, colocalization of different enzymatic activities, and increasing enzyme resilience. Here we present routes and advice for efficient design, expression, and purification of SpyTag/SpyCatcher constructs in bacterial and eukaryotic environments, including the latest 002 variants, and how to analyze reaction efficiency. The SpyInfo webpage collates the different publications and patents using SpyTag/SpyCatcher, while the SpyBank database lists their sequences and expression routes. The ability of SpyTag/SpyCatcher to react in a broad range of situations creates diverse opportunities for augmenting the function of enzymes and other biomolecules.
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spelling oxford-uuid:49069b12-afbe-46ad-972d-492a386576862022-03-26T15:29:08ZInsider information on successful covalent protein coupling with help from SpyBankBook sectionhttp://purl.org/coar/resource_type/c_3248uuid:49069b12-afbe-46ad-972d-492a38657686EnglishSymplectic Elements at OxfordElsevier2019Keeble, AHowarth, MNew biological properties can stem from the freedom to link, multimerize, or multiplex protein building blocks. The peptide SpyTag on one protein irreversibly reacts with SpyCatcher on another protein, through spontaneous isopeptide bond formation. Reaction is specific in a wide range of cellular environments and all components are genetically encoded, making this chemistry accessible to molecular biologists. SpyTag/SpyCatcher has been widely used for enzyme immobilization, colocalization of different enzymatic activities, and increasing enzyme resilience. Here we present routes and advice for efficient design, expression, and purification of SpyTag/SpyCatcher constructs in bacterial and eukaryotic environments, including the latest 002 variants, and how to analyze reaction efficiency. The SpyInfo webpage collates the different publications and patents using SpyTag/SpyCatcher, while the SpyBank database lists their sequences and expression routes. The ability of SpyTag/SpyCatcher to react in a broad range of situations creates diverse opportunities for augmenting the function of enzymes and other biomolecules.
spellingShingle Keeble, A
Howarth, M
Insider information on successful covalent protein coupling with help from SpyBank
title Insider information on successful covalent protein coupling with help from SpyBank
title_full Insider information on successful covalent protein coupling with help from SpyBank
title_fullStr Insider information on successful covalent protein coupling with help from SpyBank
title_full_unstemmed Insider information on successful covalent protein coupling with help from SpyBank
title_short Insider information on successful covalent protein coupling with help from SpyBank
title_sort insider information on successful covalent protein coupling with help from spybank
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AT howarthm insiderinformationonsuccessfulcovalentproteincouplingwithhelpfromspybank