Expanding the Scope of Sortase-Mediated Ligations by Using Sortase Homologues.

Autor: Nikghalb KD; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA., Horvath NM; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA., Prelesnik JL; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA., Banks OGB; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA., Filipov PA; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA., Row RD; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA., Roark TJ; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA., Antos JM; Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA.
Jazyk: angličtina
Zdroj: Chembiochem : a European journal of chemical biology [Chembiochem] 2018 Jan 18; Vol. 19 (2), pp. 185-195. Date of Electronic Publication: 2017 Dec 18.
DOI: 10.1002/cbic.201700517
Abstrakt: Sortase-catalyzed transacylation reactions are widely used for the construction of non-natural protein derivatives. However, the most commonly used enzyme for these strategies (sortase A from Staphylococcus aureus) is limited by its narrow substrate scope. To expand the range of substrates compatible with sortase-mediated reactions, we characterized the in vitro substrate preferences of eight sortase A homologues. From these studies, we identified sortase A enzymes that recognize multiple substrates that are unreactive toward sortase A from S. aureus. We further exploited the ability of sortase A from Streptococcus pneumoniae to recognize an LPATS substrate to perform a site-specific modification of the N-terminal serine residue in the naturally occurring antimicrobial peptide DCD-1L. Finally, we unexpectedly observed that certain substrates (LPATXG, X=Nle, Leu, Phe, Tyr) were susceptible to transacylation at alternative sites within the substrate motif, and sortase A from S. pneumoniae was capable of forming oligomers. Overall, this work provides a foundation for the further development of sortase enzymes for use in protein modification.
(© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
Databáze: MEDLINE
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