title: Sortase A as a tool to functionalize surfaces.
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.
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam, University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA Amsterdam, The Netherlands.
authors.forename: Tjitske
authors.initials: T
authors.lastname: Sijbrandij
authors.forename: Nivedita
authors.initials: N
authors.lastname: Cukkemane
authors.forename: Kamran
authors.initials: K
authors.lastname: Nazmi
authors.forename: Enno C I
authors.initials: EC
authors.lastname: Veerman
authors.forename: Floris J
authors.initials: FJ
authors.lastname: Bikker
mesh_terms: D000595:Amino Acid Sequence; D019881:Aminoacyltransferases / Q000378:metabolism*; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000378:metabolism*; D019921:Biotinylation; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000737:chemistry / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000201:enzymology*; D013499:Surface Properties; D015011:Yersinia pseudotuberculosis / Q000166:cytology
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C485059:peptide P3; D011092:Polyethylene Glycols; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases