Staphylococcus aureus Sortase A-Mediated Incorporation of Peptides: Effect of Peptide Modification on Incorporation

PloS One
Silvie Hansenová MaňáskováEnno C I Veerman

Abstract

The endogenous Staphylococcus aureus sortase A (SrtA) transpeptidase covalently anchors cell wall-anchored (CWA) proteins equipped with a specific recognition motif (LPXTG) into the peptidoglycan layer of the staphylococcal cell wall. Previous in situ experiments have shown that SrtA is also able to incorporate exogenous, fluorescently labelled, synthetic substrates equipped with the LPXTG motif (K(FITC)LPETG-amide) into the bacterial cell wall, albeit at high concentrations of 500 μM to 1 mM. In the present study, we have evaluated the effect of substrate modification on the incorporation efficiency. This revealed that (i) by elongation of LPETG-amide with a sequence of positively charged amino acids, derived from the C-terminal domain of physiological SrtA substrates, the incorporation efficiency was increased by 20-fold at 10 μM, 100 μM and 250 μM; (ii) Substituting aspartic acid (E) for methionine increased the incorporation of the resulting K(FITC)LPMTG-amide approximately three times at all concentrations tested; (iii) conjugation of the lipid II binding antibiotic vancomycin to K(FITC)LPMTG-amide resulted in the same incorporation levels as K(FITC)LPETG-amide, but much more efficient at an impressive 500-fold lower subst...Continue Reading

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Citations

Nov 18, 2017·Chemical Science·Mary J SabulskiMarcos M Pires
Dec 29, 2020·Bioorganic & Medicinal Chemistry·Jisoo ParkTae Hyeon Yoo
Jul 7, 2021·Chembiochem : a European Journal of Chemical Biology·Christine M ArtimChristopher A Alabi
Sep 8, 2021·Biotechnology and Bioengineering·Poonam KumariUtpal Mohan

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Methods Mentioned

BETA
transglycosylation
FACS
targeted modification

Software Mentioned

Prism
FacsDiva
GraphPad
GEESTNK

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