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.

Simple Method of Synthesizing Nickel–Nitrilotriacetic Acid Gold Nanoparticles with a Narrow Size Distribution for Protein Labeling

“…Various ligands, such as coenzyme A (CoA) derivatives, 11) benzylguanine derivatives, 12) and benzylcytosine derivatives, 13) have been developed to label biomolecules. 14,15) The functionalization of Au NPs with thiol-containing derivatives of such ligands should also be valuable in identifying specific sites in proteins.…”

Tension Monitoring during Epithelial-to-Mesenchymal Transition Links the Switch of Phenotype to Expression of Moesin and Cadherins in NMuMG Cells

“…This coupling is induced via a Snap-Tag C-terminally fused to Ecad15. The coupling procedure is described in detail by Wedlich and coworkers [36]. Force-distance measurements were performed using a pulling speed of 1 µm/s and a contact force of 100 pN.…”

Dynamic Covalent Chemistry on Surfaces Employing Highly Reactive Cyclopentadienyl Moieties

“…[ 17 ] The C 1s signal is additionally visible and can be deconvoluted into three peaks centered at 285.0, 286.3, and 288.3 eV corresponding to the C-C/C-H, C-N, and C = O structural features, respectively. [ 16 ] On inspection of this peak it is noted that the total atom percentage contribution from the C-N signal is slightly Figure S4 (Supporting Information). In order to estimate the layers' thickness, an area of the polysiloxane layer was removed from the silicon wafer by scratching with an AFM cantilever, leaving the bare wafer exposed.…”

“…In particular, the N 1s signal, which appears at 400.1 eV and is associated with the amide linkage, [ 16 ] and the Br 3d signal at 70.3 eV due to the terminal bromide are visible. [ 17 ] The C 1s signal is additionally visible and can be deconvoluted into three peaks centered at 285.0, 286.3, and 288.3 eV corresponding to the C-C/C-H, C-N, and C = O structural features, respectively.…”