A Versatile Toolbox for Multiplexed Protein Micropatterning by Laser Lithography

Autor(en): Gropeanu, Mihaela
Bhagawati, Maniraj
Gropeanu, Radu A.
Muniz, Gemma M. Rodriguez
Sundaram, Subramanian
Piehler, Jacob 
del Campo, Aranzazu
Stichwörter: BIOMOLECULES; Chemistry; Chemistry, Multidisciplinary; Chemistry, Physical; FABRICATION; FUNCTIONAL IMMOBILIZATION; GLASS; histidine tags; HISTIDINE-TAGGED PROTEINS; laser lithography; Materials Science; Materials Science, Multidisciplinary; MOTOR PROTEINS; Nanoscience & Nanotechnology; photocleavable peptides; PHOTOLABILE PROTECTING GROUPS; phototriggers; Physics; Physics, Applied; Physics, Condensed Matter; protein micropatterns; RECEPTOR INTERACTIONS; Science & Technology - Other Topics; SURFACE; THIOLS
Erscheinungsdatum: 2013
Journal: SMALL
Volumen: 9
Ausgabe: 6
Startseite: 838
Seitenende: 845
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.
ISSN: 16136810
DOI: 10.1002/smll.201201901

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