Native laser lithography of His-tagged proteins by uncaging of multivalent chelators.
We report a generic approach for targeting proteins into micropatterns by in situ laser lithography. To this end, we have designed a photocleavable oligohistidine peptide for caging tris(nitrilo triacetic acid) (tris-NTA) groups on surfaces by multivalent interactions. Local photofragmentation of the peptide by UV illumination through a photomask or by a confocal laser beam uncages tris-NTA, thus generating free binding sites for rapid, site-specific capturing of His-tagged proteins into micropatterns. Iterative writing of proteins by laser lithography enabled for assembly of multiplexed functional protein microstructures on surfaces. Thus, versatile, user-defined protein micropatterns can be assembled under physiological conditions with a standard confocal laser-scanning microscope.
Bhagawati
Maniraj
M
Division of Biophysics, University of Osnabrück, 49076 Osnabrück, Germany.
Lata
Suman
S
Tampé
Robert
R
Piehler
Jacob
J
D002614:Chelating Agents / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D007834:Lasers; D049329:Nanostructures / Q000737:chemistry*; D009571:Nitrilotriacetic Acid / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D011327:Printing / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics; D014466:Ultraviolet Rays
D002614:Chelating Agents; D010455:Peptides; D011994:Recombinant Proteins; D006639:Histidine; D009571:Nitrilotriacetic Acid