title: Tyrosinase-catalyzed site-specific immobilization of engineered C-phycocyanin to surface.
abstract: Enzymatic crosslinking of proteins is often limited by the steric availability of the target residues, as of tyrosyl side chains in the case of tyrosinase. Carrying an N-terminal peptide-tag containing two tyrosine residues, the fluorescent protein C-phycocyanin HisCPC from Synechocystis sp. PCC6803 was crosslinked to fluorescent high-molecular weight forms with tyrosinase. Crosslinking with tyrosinase in the presence of L-tyrosine produced non fluorescent high-molecular weight products. Incubated in the presence of tyrosinase, HisCPC could also be immobilized to amino-modified polystyrene beads thus conferring a blue fluorescence. Crosslinking and immobilization were site-specific as both processes required the presence of the N-terminal peptide in HisCPC.
authors.affiliation: Empa, Swiss Federal Laboratories for Materials Science and Technology - Laboratory for Bioactive Materials, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland.
authors.forename: Greta
authors.initials: G
authors.lastname: Faccio
authors.affiliation: Empa, Swiss Federal Laboratories for Materials Science and Technology - Laboratory for Bioactive Materials, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Kämpf
authors.affiliation: 1] Empa, Swiss Federal Laboratories for Materials Science and Technology - Laboratory for Bioactive Materials, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland [2] Dipartimento di Biotecnologie e Scienze della Vita, Università degli Studi dell'Insubria, via J.H. Dunant 3, 21100 Varese, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Piatti
authors.affiliation: Empa, Swiss Federal Laboratories for Materials Science and Technology - Laboratory for Bioactive Materials, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland.
authors.forename: Linda
authors.initials: L
authors.lastname: Thöny-Meyer
authors.affiliation: Empa, Swiss Federal Laboratories for Materials Science and Technology - Laboratory for Bioactive Materials, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Richter
mesh_terms: D001665:Binding Sites; D002384:Catalysis; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004355:Drug Stability; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005557:Formaldehyde / Q000737:chemistry*; D014442:Monophenol Monooxygenase / Q000737:chemistry*; D010798:Phycocyanin / Q000737:chemistry*; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry; D046939:Synechocystis / Q000201:enzymology* / Q000235:genetics
chemical_list: D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D010798:Phycocyanin; D005557:Formaldehyde; D014442:Monophenol Monooxygenase