title: Site-specific protein immobilization using unnatural amino acids.
abstract: Protein immobilization confers the advantages of biological systems to a more chemical setting and has applications in catalysis, sensors, and materials development. While numerous immobilization techniques exist, it is optimal to develop a well-defined and chemically stable methodology to allow for full protein function. This paper describes the utilization of unnatural amino acid technologies to introduce bioorthogonal handles in a site-specific fashion for protein immobilization. To develop this approach a range of solid-supports, organic linkers, and protein immobilization sites have been investigated using a GFP reporter system. Overall, a sepharose resin derivatized with propargyl alcohol has afforded the highest yields of immobilized protein. Moreover, an unnatural amino acid residue protein context has been demonstrated, signifying a necessity to consider the protein site of immobilization. Finally, a resin-conferred stabilization was demonstrated in several organic solvents.
authors.affiliation: Department of Chemistry, College of William & Mary P.O. Box 8795, Williamsburg, Virginia 23187, United States.
authors.forename: Benjamin K
authors.initials: BK
authors.lastname: Raliski
authors.forename: Christina A
authors.initials: CA
authors.lastname: Howard
authors.forename: Douglas D
authors.initials: DD
authors.lastname: Young
mesh_terms: D000480:Alkynes / Q000737:chemistry; D000596:Amino Acids / Q000737:chemistry*; D001665:Binding Sites; D055767:Immobilized Proteins / Q000737:chemistry*; D008958:Models, Molecular; D020005:Propanols / Q000737:chemistry; D017433:Protein Structure, Secondary; D012685:Sepharose / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013379:Substrate Specificity
chemical_list: D000480:Alkynes; D000596:Amino Acids; D055767:Immobilized Proteins; D020005:Propanols; D012997:Solvents; D012685:Sepharose; C028255:propargyl alcohol