Functional glass surface displaying a glutamyl donor substrate for transglutaminase-mediated protein immobilization.
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.
Sung
Kyunga
K
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.
Kamiya
Noriho
N
Kawata
Noriyuki
N
Kamiya
Shinji
S
Goto
Masahiro
M
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*
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