Functional Immobilization and Patterning of Proteins by an Enzymatic Transfer Reaction
DC Element | Wert | Sprache |
---|---|---|
dc.contributor.author | Waichman, Sharon | |
dc.contributor.author | Bhagawati, Maniraj | |
dc.contributor.author | Podoplelova, Yulia | |
dc.contributor.author | Reichel, Annett | |
dc.contributor.author | Brunk, Ariane | |
dc.contributor.author | Paterok, Dirk | |
dc.contributor.author | Piehler, Jacob | |
dc.date.accessioned | 2021-12-23T16:03:51Z | - |
dc.date.available | 2021-12-23T16:03:51Z | - |
dc.date.issued | 2010 | |
dc.identifier.issn | 00032700 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/6233 | - |
dc.description.abstract | Functional immobilization and lateral organization of proteins into micro- and nanopatterns is an important prerequisite for miniaturizing bioanalytical and biotechnological devices. Here, we report an approach for efficient site-specific protein immobilization based on enzymatic phosphopantetheinyl transfer (PPT) from coenzyme A (CoA)-functionalized glass-type surfaces to specific peptide tags. We devised a bottom-up surface modification approach for coupling CoA densely to a molecular poly(ethylene glycol) polymer brush. Site-specific enzymatic immobilization of proteins fused to different target peptides for the PPTase Sfp was confirmed by real-time label-free detection. Quantitative protein-protein interaction experiments confirmed that significantly more than 50% of the immobilized protein was fully active. The method was successfully applied with different proteins. However, different immobilization efficiencies of PPT-based immobilization were observed for different peptide tags being fused to the N- and C-termini of proteins. On the basis of this immobilization method, we established photolithographic patterning of proteins into functional binary microstructures. | |
dc.description.sponsorship | DFGGerman Research Foundation (DFG)European Commission [PI 405-4, EXC 115, PI 405-3]; BMBFFederal Ministry of Education & Research (BMBF) [0312034]; Minerva Foundation; We thank Gabriele Hikade and Hella Kenneweg for protein production, Covalys Biosciences for technical support with PPT technology, and NB-Technologies for providing photomasks. Ibis project was supported by funding from the DFG (PI 405-4 and EXC 115) and by the BMBF (0312034). J.P was supported by a Heisenberg Professorship from the DFG (PI 405-3) and S.W. 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 INTERFACE | |
dc.subject | Chemistry | |
dc.subject | Chemistry, Analytical | |
dc.subject | FUSION PROTEINS | |
dc.subject | HISTIDINE-TAGGED PROTEINS | |
dc.subject | I INTERFERON-RECEPTOR | |
dc.subject | IFNAR1 | |
dc.subject | MOTOR PROTEINS | |
dc.subject | SFP PHOSPHOPANTETHEINYL TRANSFERASE | |
dc.subject | SMALL MOLECULES | |
dc.subject | SURFACES | |
dc.subject | TECHNOLOGIES | |
dc.title | Functional Immobilization and Patterning of Proteins by an Enzymatic Transfer Reaction | |
dc.type | journal article | |
dc.identifier.doi | 10.1021/ac902608a | |
dc.identifier.isi | ISI:000274466100044 | |
dc.description.volume | 82 | |
dc.description.issue | 4 | |
dc.description.startpage | 1478 | |
dc.description.endpage | 1485 | |
dc.identifier.eissn | 15206882 | |
dc.publisher.place | 1155 16TH ST, NW, WASHINGTON, DC 20036 USA | |
dcterms.isPartOf.abbreviation | Anal. Chem. | |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.orcid | 0000-0002-2143-2270 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | PiJa938 | - |
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geprüft am 02.05.2024