Investigation of selective protein immobilization on charged protein array by wavelength interrogation-based SPR sensor.
We have investigated the use of multilayer films of polyelectrolytes as selective surfaces to analyze protein interactions with a self-assembled SPR wavelength-shift sensor. Charged arrays were prepared by alternating adsorption of the charged polyelectrolytes, poly(diallyldimethylammonium chloride) (PDDA) and poly(sodium 4-styrenesulfonate) (PSS). Multilayer formation was monitored with the SPR wavelength-shift sensor and a Spreeta SPR sensor. Protein immobilization on the charged surfaces, which was also analyzed by the SPR sensors, was dependent on the pI of the proteins. Tissue transglutaminase (tTGase) and beta-galactosidase (pIs, 5.1 and 5.3, respectively) were preferentially bound to the positively charged PDDA surface, whereas lysozyme (pI, 11.0) was selectively bound to the negatively charged PSS surface. Immobilization of tTGase on the PDDA surface was also dependent on the buffer pH. The interaction of tTGase with RhoA(V14), a constitutively active form of Rho, could be detected on the charged arrays with the wavelength-shift sensor. The arrays could be reutilized at least 5 times. Thus, it is likely that charged surfaces, assembled by the layer-by-layer method using polyelectrolytes, will prove useful for preparing selective protein arrays.
Yi
Sun-Ju
SJ
Department of Molecular and Cellullar Biochemistry, Kangwon National University School of Medicine, Chunchon 200-701, Korea.
Yuk
Jong Seol
JS
Jung
Se-Hui
SH
Zhavnerko
Genady K
GK
Kim
Young-Myeong
YM
Ha
Kwon-Soo
KS
D000327:Adsorption; D008689:Methacrylates; D009113:Muramidase / Q000378:metabolism; D011095:Polyethylenes; D011108:Polymers; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011725:Pyridines; D000644:Quaternary Ammonium Compounds; D013451:Sulfonic Acids; D020349:Surface Plasmon Resonance; D011503:Transglutaminases / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism; D020742:rhoA GTP-Binding Protein
D008689:Methacrylates; D011095:Polyethylenes; D011108:Polymers; D011725:Pyridines; D000644:Quaternary Ammonium Compounds; D013451:Sulfonic Acids; C400644:poly(diphenyl-2-pyridylmethyl methacrylate); C041004:poly-N,N-dimethyl-N,N-diallylammonium chloride; D011503:Transglutaminases; D009113:Muramidase; D001616:beta-Galactosidase; D020742:rhoA GTP-Binding Protein; C077114:styrenesulfonic acid polymer