posted on 2019-09-12, 12:35authored byMohammad
A. Rahman, Cordula Reuther, Frida W. Lindberg, Martina Mengoni, Aseem Salhotra, Georg Heldt, Heiner Linke, Stefan Diez, Alf Månsson
The
guided gliding of cytoskeletal filaments, driven by biomolecular
motors on nano/microstructured chips, enables novel applications in
biosensing and biocomputation. However, expensive and time-consuming
chip production hampers the developments. It is therefore important
to establish protocols to regenerate the chips, preferably without
the need to dismantle the assembled microfluidic devices which contain
the structured chips. We here describe a novel method toward this
end. Specifically, we use the small, nonselective proteolytic enzyme,
proteinase K to cleave all surface-adsorbed proteins, including myosin
and kinesin motors. Subsequently, we apply a detergent (5% SDS or
0.05% Triton X100) to remove the protein remnants. After this procedure,
fresh motor proteins and filaments can be added for new experiments.
Both, silanized glass surfaces for actin–myosin motility and
pure glass surfaces for microtubule–kinesin motility were repeatedly
regenerated using this approach. Moreover, we demonstrate the applicability
of the method for the regeneration of nano/microstructured silicon-based
chips with selectively functionalized areas for supporting or suppressing
gliding motility for both motor systems. The results substantiate
the versatility and a promising broad use of the method for regenerating
a wide range of protein-based nano/microdevices.