A chemically modified glass surface that facilitates transglutaminase-mediated protein immobilization.
An amino-modified glass surface for enzymatic protein immobilization by microbial transglutaminase (MTG) was developed. Diamine substrates with secondary amino groups in the linker moiety, like triethylenetetramine (TETA), exhibited at most a 2-fold higher reactivity in the MTG-catalyzed reaction compared to those with the alkyl linker. A 96-well glass plate was subsequently modified with selected diamine substrates. Validation of the modified surface by enzymatic immobilization of enhanced green fluorescent protein tagged with a glutamine donor-substrate peptide (LLQG) of MTG revealed that the protein loading onto the TETA-modified glass surface was approximately 15-fold higher than that on the unmodified one.
Tanaka
Yusuke
Y
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Fukuoka, 819-0395, Japan.
Doi
Satoshi
S
Kamiya
Noriho
N
Kawata
Noriyuki
N
Kamiya
Shinji
S
Nakama
Kenichi
K
Goto
Masahiro
M
D001426:Bacterial Proteins / Q000737:chemistry; D002384:Catalysis; D005898:Glass / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D013499:Surface Properties; D011503:Transglutaminases / Q000378:metabolism*; D014266:Trientine / Q000737:chemistry*
D001426:Bacterial Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011503:Transglutaminases; D014266:Trientine