title: Benzylguanine thiol self-assembled monolayers for the immobilization of SNAP-tag proteins on microcontact-printed surface structures.
abstract: The site-selective, oriented, covalent immobilization of proteins on surfaces is an important issue in the establishment of microarrays, biosensors, biocatalysts, and cell assays. Here we describe the preparation of self-assembled monolayers consisting of benzylguanine thiols (BGT) to which SNAP-tag fusion proteins can be covalently linked. The SNAP-tag, a modified O(6)-alkylguanine-DNA alkyltransferase (AGT), reacts with the headgroup of BGT and becomes covalently bound upon the release of guanine. Bacterially produced recombinant His-tag-SNAP-tag-GFP was used to demonstrate the site-specific immobilization on BGT surface patterns created by microcontact printing (microCP). With this versatile method, any SNAP-tag protein can be coupled to a surface.
authors.affiliation: Center for Functional Nanostructures, Karlsruhe Institute for Technology, Kaiserstrasse 12, D-76131 Karlsruhe, Germany.
authors.forename: Sinem
authors.initials: S
authors.lastname: Engin
authors.forename: Vanessa
authors.initials: V
authors.lastname: Trouillet
authors.forename: Clemens M
authors.initials: CM
authors.lastname: Franz
authors.forename: Alexander
authors.initials: A
authors.lastname: Welle
authors.forename: Michael
authors.initials: M
authors.lastname: Bruns
authors.forename: Doris
authors.initials: D
authors.lastname: Wedlich
mesh_terms: D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D006146:Guanidines / Q000737:chemistry*; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000737:chemistry* / Q000378:metabolism; D011327:Printing; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D013499:Surface Properties
chemical_list: D004800:Enzymes, Immobilized; D006146:Guanidines; D011993:Recombinant Fusion Proteins; D019853:O(6)-Methylguanine-DNA Methyltransferase