In situ regeneration of bioactive coatings enabled by an evolved Staphylococcus aureus sortase A

Nat Commun. 2016 Apr 13;7:11140. doi: 10.1038/ncomms11140.

Abstract

Surface immobilization of bioactive molecules is a central paradigm in the design of implantable devices and biosensors with improved clinical performance capabilities. However, in vivo degradation or denaturation of surface constituents often limits the long-term performance of bioactive films. Here we demonstrate the capacity to repeatedly regenerate a covalently immobilized monomolecular thin film of bioactive molecules through a two-step stripping and recharging cycle. Reversible transpeptidation by a laboratory evolved Staphylococcus aureus sortase A (eSrtA) enabled the rapid immobilization of an anti-thrombogenic film in the presence of whole blood and permitted multiple cycles of film regeneration in vitro that preserved its biological activity. Moreover, eSrtA transpeptidation facilitated surface re-engineering of medical devices in situ after in vivo implantation through removal and restoration film constituents. These studies establish a rapid, orthogonal and reversible biochemical scheme to regenerate selective molecular constituents with the potential to extend the lifetime of bioactive films.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoacyltransferases / metabolism*
  • Animals
  • Bacterial Proteins / metabolism*
  • Biocatalysis / drug effects
  • Catheterization, Peripheral
  • Coated Materials, Biocompatible / pharmacology*
  • Cysteine Endopeptidases / metabolism*
  • Mice, Inbred C57BL
  • Rats, Wistar
  • Staphylococcus aureus / enzymology*
  • Surface Properties

Substances

  • Bacterial Proteins
  • Coated Materials, Biocompatible
  • Aminoacyltransferases
  • sortase A
  • Cysteine Endopeptidases