Research Mentor(s)
Antos, John M.
Description
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.
Document Type
Event
Start Date
18-5-2017 9:00 AM
End Date
18-5-2017 12:00 PM
Department
Chemistry
Genre/Form
student projects; posters
Subjects – Topical (LCSH)
Enzymes--Synthesis--Research; Protein engineering; Streptococcus pneumoniae--Research
Type
Image
Rights
Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this documentation for commercial purposes, or for financial gain, shall not be allowed without the author's written permission.
Language
English
Format
application/pdf
Included in
Broadening the Scope of Sortase-Mediated Ligations using Natural Sortase Homologs
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.