Sortase A as a tool to functionalize surfaces

Bioconjug Chem. 2013 May 15;24(5):828-31. doi: 10.1021/bc4000146. Epub 2013 Apr 10.

Abstract

A widely accepted approach to combat surface fouling is based on the prevention of biofoulants to attach to a surface by the functionalization with poly(ethylene glycol) (PEG). The goal of this study was to generate a proof of concept for the enzymatic coupling of PEG to a peptide precoated surface by using the enzyme Sortase A (SrtA). A hydrophobic polystyrene surface was primed with anchoring peptide P3 equipped with a pentaglycine acceptor motif for SrtA, to enable subsequent transpeptidation with either biotin or a PEG-tail containing the sortase recognition motif LPETG. High levels of surface-bound biotin were detected only in cases with biotin-LPETG and SrtA. Little if any reactivity was detected in wells treated with the SrtA scrambled motif EGLTP, or in the absence of SrtA. Conjugation of PEG resulted in a significant decrease of bacterial adherence to the surface.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aminoacyltransferases / metabolism*
  • Bacterial Adhesion
  • Bacterial Proteins / metabolism*
  • Biotinylation
  • Carrier Proteins / chemistry
  • Carrier Proteins / metabolism*
  • Cysteine Endopeptidases / metabolism*
  • Humans
  • Intercellular Signaling Peptides and Proteins
  • Peptides / chemistry
  • Peptides / metabolism*
  • Polyethylene Glycols / chemistry
  • Polyethylene Glycols / metabolism*
  • Staphylococcus aureus / enzymology*
  • Surface Properties
  • Yersinia pseudotuberculosis / cytology

Substances

  • Bacterial Proteins
  • Carrier Proteins
  • Intercellular Signaling Peptides and Proteins
  • Peptides
  • peptide P3
  • Polyethylene Glycols
  • Aminoacyltransferases
  • sortase A
  • Cysteine Endopeptidases