title: Covalent attachment of proteins to solid supports and surfaces via Sortase-mediated ligation.
abstract: BACKGROUND
  There is growing interest in the attachment of proteins to solid supports for the development of supported catalysts, affinity matrices, and micro devices as well as for the development of planar and bead based protein arrays for multiplexed assays of protein concentration, interactions, and activity. A critical requirement for these applications is the generation of a stable linkage between the solid support and the immobilized, but still functional, protein.
  
  
  METHODS
  Solid supports including crosslinked polymer beads, beaded agarose, and planar glass surfaces, were modified to present an oligoglycine motif to solution. A range of proteins were ligated to the various surfaces using the Sortase A enzyme of S. aureus. Reactions were carried out in aqueous buffer conditions at room temperature for times between one and twelve hours.
  
  
  CONCLUSIONS
  The Sortase A transpeptidase of S. aureus provides a general, robust, and gentle approach to the selective covalent immobilization of proteins on three very different solid supports. The proteins remain functional and accessible to solution. Sortase mediated ligation is therefore a straightforward methodology for the preparation of solid supported enzymes and bead based assays, as well as the modification of planar surfaces for microanalytical devices and protein arrays.
authors.affiliation: School of Chemistry, University of Southampton, Highfield, Southampton, United Kingdom.
authors.forename: Lilyan
authors.initials: L
authors.lastname: Chan
authors.forename: Hannah F
authors.initials: HF
authors.lastname: Cross
authors.forename: Joseph K
authors.initials: JK
authors.lastname: She
authors.forename: Gabriel
authors.initials: G
authors.lastname: Cavalli
authors.forename: Hugo F P
authors.initials: HF
authors.lastname: Martins
authors.forename: Cameron
authors.initials: C
authors.lastname: Neylon
mesh_terms: D001483:Base Sequence; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D017931:DNA Primers; D006868:Hydrolysis; D008854:Microscopy, Electron; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D011506:Proteins; D003546:Cysteine Endopeptidases