DC Field | Value | Language |
dc.contributor.author | Waichman, Sharon | |
dc.contributor.author | You, Changjiang | |
dc.contributor.author | Beutel, Oliver | |
dc.contributor.author | Bhagawati, Maniraj | |
dc.contributor.author | Piehler, Jacob | |
dc.date.accessioned | 2021-12-23T16:04:25Z | - |
dc.date.available | 2021-12-23T16:04:25Z | - |
dc.date.issued | 2011 | |
dc.identifier.issn | 00032700 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/6416 | - |
dc.description.abstract | Spatial organization of proteins into microscopic structures has important applications in fundamental and applied research. Preserving the function of proteins in such microstructures requires generic methods for site-specific capturing through affinity handles. Here, we present a versatile bottom-up surface micropatterning approach based on surface functionalization with male-imides, which selectively react with organic thiols. Upon UV irradiation through a photomask, the functionality of illuminated maleimide groups was efficiently destroyed. Remaining maleimides in nonilluminated regions were further reacted with different thiol-functionalized groups for site-specific protein immobilization under physiological conditions. Highly selective immobilization of His-tagged proteins into tris(nitrilotriacetic acid) functionalized microstructures with very high contrast was possible even by direct capturing of proteins from crude cell lysates. Moreover, we employed phosphopantetheinyl transfer from surface-immobilized coenzyme A to ybbR-tagged proteins in order to implement site-specific, covalent protein immobilization into microstructures. The functional integrity of the immobilized protein was confirmed by monitoring protein-protein interactions in real time. Moreover, we demonstrate quantitative single-molecule analysis of protein-protein interactions with proteins selectively captured into these high-contrast micropatterns. | |
dc.description.sponsorship | BMBFFederal Ministry of Education & Research (BMBF) [0312034]; DFGGerman Research Foundation (DFG)European Commission [PI 405/3, PI 405/4]; Minerva Foundation; We thank Gabriele Hikade and Hella Kenneweg for technical assistance and NB Technologies for providing photomasks. This project was supported by the BMBF (0312034) and by the DFG (PI 405/3, PI 405/4). S.W. was supported by a Ph.D. fellowship from the Minerva Foundation. | |
dc.language.iso | en | |
dc.publisher | AMER CHEMICAL SOC | |
dc.relation.ispartof | ANALYTICAL CHEMISTRY | |
dc.subject | BINDING | |
dc.subject | BIOTIN | |
dc.subject | Chemistry | |
dc.subject | Chemistry, Analytical | |
dc.subject | FUNCTIONAL IMMOBILIZATION | |
dc.subject | HISTIDINE-TAGGED PROTEINS | |
dc.subject | MOTOR PROTEINS | |
dc.subject | RECEPTOR INTERACTIONS | |
dc.subject | REFLECTOMETRIC INTERFERENCE SPECTROSCOPY | |
dc.subject | SFP PHOSPHOPANTETHEINYL TRANSFERASE | |
dc.subject | STREPTAVIDIN | |
dc.subject | SURFACE | |
dc.title | Maleimide Photolithography for Single-Molecule Protein-Protein Interaction Analysis in Micropatterns | |
dc.type | journal article | |
dc.identifier.doi | 10.1021/ac1021453 | |
dc.identifier.isi | ISI:000286129800006 | |
dc.description.volume | 83 | |
dc.description.issue | 2 | |
dc.description.startpage | 501 | |
dc.description.endpage | 508 | |
dc.contributor.orcid | 0000-0002-7839-6397 | |
dc.contributor.researcherid | L-3901-2014 | |
dc.identifier.eissn | 15206882 | |
dc.publisher.place | 1155 16TH ST, NW, WASHINGTON, DC 20036 USA | |
dcterms.isPartOf.abbreviation | Anal. Chem. | |
crisitem.author.dept | Sonderforschungsbereich 944: Physiologie und Dynamik zellulärer Mikrokompartimente | - |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | organisation19 | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.orcid | 0000-0002-7839-6397 | - |
crisitem.author.orcid | 0000-0002-2143-2270 | - |
crisitem.author.parentorg | FB 05 - Biologie/Chemie | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.grandparentorg | Universität Osnabrück | - |
crisitem.author.netid | YoCh745 | - |
crisitem.author.netid | PiJa938 | - |