Research Mentor(s)

John Antos

Affiliated Department

Chemistry

Sort Order

12

Start Date

18-5-2017 9:00 AM

End Date

18-5-2017 12:00 PM

Document Type

Event

Abstract

Sortase-mediated ligations have become an attractive option for protein modification chemistry, enabling the synthesis of a wide range of non-natural polypeptide derivatives. In an effort to expand the scope of this methodology, we have been characterizing the in vitro reactivity of a panel of natural sortase homologs. Here we present our studies on the substrate and nucleophile tolerance of sortases from a range of bacterial species. Notable findings include that sortase A from Streptococcus pneumoniae (SrtApneu) shows a high degree of substrate promiscuity, allowing this enzyme to process a range of substrate variations that deviate from the LPXTG substrate motif typically associated with sortase-mediated methods. In addition, this enzyme has the ability to accept an expanded range of primary amine nucleophiles. To demonstrate the utility of this expanded substrate scope, we have also succeeded in using SrtApneu to site-specifically modify the N-terminal serine residue of Dermcidin (DCD-1L). Overall, these results demonstrate that naturally occurring sortases represent a viable approach for the continued development of sortase-mediated protein modification.

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Language

English

Format

application/pdf

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May 18th, 9:00 AM May 18th, 12:00 PM

Broadening the Scope of Sortase-Mediated Ligations using Natural Sortase Homologs

Chemistry

Sortase-mediated ligations have become an attractive option for protein modification chemistry, enabling the synthesis of a wide range of non-natural polypeptide derivatives. In an effort to expand the scope of this methodology, we have been characterizing the in vitro reactivity of a panel of natural sortase homologs. Here we present our studies on the substrate and nucleophile tolerance of sortases from a range of bacterial species. Notable findings include that sortase A from Streptococcus pneumoniae (SrtApneu) shows a high degree of substrate promiscuity, allowing this enzyme to process a range of substrate variations that deviate from the LPXTG substrate motif typically associated with sortase-mediated methods. In addition, this enzyme has the ability to accept an expanded range of primary amine nucleophiles. To demonstrate the utility of this expanded substrate scope, we have also succeeded in using SrtApneu to site-specifically modify the N-terminal serine residue of Dermcidin (DCD-1L). Overall, these results demonstrate that naturally occurring sortases represent a viable approach for the continued development of sortase-mediated protein modification.

 

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