title: A versatile toolbox for multiplexed protein micropatterning by laser lithography.
abstract: Photocleavable oligohistidine peptides (POHP) allow in situ spatial organization of multiple His-tagged proteins onto surfaces functionalized with tris(nitrilotriacetic acid) (tris-NTA). Here, a second generation of POHPs is presented with improved photoresponse and site-specific covalent coupling is introduced for generating stable protein assemblies. POHPs with different numbers of histidine residues and a photocleavable linker based on the 4,5-dimethoxy-o-nitrophenyl ethyl chromophore are prepared. These peptides show better photosensitivity than the previously used o-nitrophenyl ethyl derivative. Efficient and stable caging of tris-NTA-functionalized surfaces by POHPs comprising 12 histidine residues is demonstrated by multiparameter solid-phase detection techniques. Laser lithographic uncaging by photofragmentation of the POHPs is possible with substantially reduced photodamage as compared to previous approaches. Thus, in situ micropatterning of His-tagged proteins under physiological conditions is demonstrated for the first time. In combination with a short peptide tag for a site-specific enzymatic coupling reaction, covalent immobilization of multiple proteins into target micropatterns is possible under physiological conditions.
authors.affiliation: Max-Planck-Institut für Polymerforschung, Mainz, Germany.
authors.forename: Mihaela
authors.initials: M
authors.lastname: Gropeanu
authors.forename: Maniraj
authors.initials: M
authors.lastname: Bhagawati
authors.forename: Radu A
authors.initials: RA
authors.lastname: Gropeanu
authors.forename: Gemma M
authors.initials: GM
authors.lastname: Rodríguez Muñiz
authors.forename: Subramanian
authors.initials: S
authors.lastname: Sundaram
authors.forename: Jacob
authors.initials: J
authors.lastname: Piehler
authors.forename: Aránzazu
authors.initials: A
authors.lastname: del Campo
mesh_terms: D006639:Histidine / Q000737:chemistry; D007834:Lasers; D055668:Photochemical Processes; D011506:Proteins / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D011506:Proteins; D006639:Histidine