Characterization of covalently bonded proteins on poly(methyl methacrylate) by X-ray photoelectron spectroscopy.

Autor: Nelson GW; Department of Chemistry, Queen's University, Kingston, Ontario, Canada., Perry M, He SM, Zechel DL, Horton JH
Jazyk: angličtina
Zdroj: Colloids and surfaces. B, Biointerfaces [Colloids Surf B Biointerfaces] 2010 Jun 15; Vol. 78 (1), pp. 61-8. Date of Electronic Publication: 2010 Feb 18.
DOI: 10.1016/j.colsurfb.2010.02.012
Abstrakt: X-ray photoelectron spectroscopy (XPS) has been used to characterize a poly(methyl methacrylate) (PMMA) surface with covalently attached proteins. The PMMA surfaces were first aminated using hexamethyldiamine; the resulting -NH(2) sites were reacted with the hetero-bifunctional cross-linker Sulfo-EMCS to form a maleimide-terminated surface. The N-hydroxysuccinimide ester terminal and maleimide terminal groups of Sulfo-EMCS reacts with amine and sulfhydryl groups, respectively, exposed on the surface of the proteins. This study characterizes Thermotoga maritima beta-glucosidase 1 (TmGH1), which belongs to a family of proteins that facilitate hydrolysis of glucose-related monomers with retention of conformation. The surfaces were characterized by XPS to monitor surface composition, and to elucidate protein orientation on the surface. Results suggest that a covalently bonded surface of TmGH1 on PMMA has been obtained. These results demonstrate the feasibility of using XPS to study protein surface chemistry and demonstrate a useful method to anchor cysteine-terminated proteins for the purposes of creating biosensors or platforms for mechanical force experiments to investigate protein structure.
(Copyright 2010 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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