Functional modification of fibronectin by N-terminal FXIIIa-mediated transamidation.

Autor: Früh SM; Department of Health Sciences and Technology, ETH Zürich, Vladimir-Prelog-Weg 4, 8093 Zürich (Switzerland) http://www.appliedmechanobio.ethz.ch., Spycher PR, Mitsi M, Burkhardt MA, Vogel V, Schoen I
Jazyk: angličtina
Zdroj: Chembiochem : a European journal of chemical biology [Chembiochem] 2014 Jul 07; Vol. 15 (10), pp. 1481-6. Date of Electronic Publication: 2014 Jun 06.
DOI: 10.1002/cbic.201402099
Abstrakt: A straightforward strategy is presented for the site-specific incorporation of fluorophores or reactive probes into the extracellular matrix (ECM) protein fibronectin (Fn) by using the enzyme-catalyzed transamidation by activated factor XIII. Characterization by SDS-PAGE, western blotting, absorption measurements, mass spectrometry, and stepwise photobleaching for labeling quantification at the single-molecule level showed that the labeling was efficient and restricted to the N-terminal tails. The introduction of labels did not interfere with Fn fibrillogenesis, as verified by the incorporation of fluorescently labeled Fn into ECM and manually pulled Fn fibers. Site-specific incorporation of an azide was used to create a template for bioorthogonal click chemistry reactions in a second bioconjugation step, thus offering versatile modification and application possibilities in the context of matrix biology and tissue engineering.
(© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
Databáze: MEDLINE
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