Benzylguanine thiol self-assembled monolayers for the immobilization of SNAP-tag proteins on microcontact-printed surface structures.
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.
Engin
Sinem
S
Center for Functional Nanostructures, Karlsruhe Institute for Technology, Kaiserstrasse 12, D-76131 Karlsruhe, Germany.
Trouillet
Vanessa
V
Franz
Clemens M
CM
Welle
Alexander
A
Bruns
Michael
M
Wedlich
Doris
D
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
D004800:Enzymes, Immobilized; D006146:Guanidines; D011993:Recombinant Fusion Proteins; D019853:O(6)-Methylguanine-DNA Methyltransferase