title: Functional glass surface displaying a glutamyl donor substrate for transglutaminase-mediated protein immobilization.
abstract: A chemically modified glass surface displaying a glutamyl donor substrate peptide (Z-QG) was developed for microbial transglutaminase (MTG)-mediated immobilization of recombinant proteins tagged with an MTG-reactive lysine-containing substrate peptide (K-tag). To evaluate the surface modification conditions affecting the enzymatic protein immobilization, we employed an amino-modified 96-well glass plate as a base and prepared three types of glass surfaces displaying Z-QG. Validation of the Z-QG modified glass surfaces with recombinant enhanced green fluorescent proteins revealed that the insertion of a di(ethylene glycol) linker between the terminal Z-QG moiety and the base not only enhances enzymatic protein immobilization efficiency but also decreases nonselective protein adsorption. A bacterial alkaline phosphatase fused with a K-tag at the N terminus was also successfully immobilized to the designed glass surface, suggesting that the chemically modified glass surface displaying a spatially controlled glutamyl donor substrate is a potential platform for MTG-mediated fabrication of protein-based solid biomaterials.
authors.affiliation: Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.
authors.forename: Kyunga
authors.initials: K
authors.lastname: Sung
authors.forename: Noriho
authors.initials: N
authors.lastname: Kamiya
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kawata
authors.forename: Shinji
authors.initials: S
authors.lastname: Kamiya
authors.forename: Masahiro
authors.initials: M
authors.lastname: Goto
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D005898:Glass; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D055767:Immobilized Proteins; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011503:Transglutaminases; D000469:Alkaline Phosphatase