A Versatile Toolbox for Multiplexed Protein Micropatterning by Laser Lithography

作者：Gropeanu Mihaela; Bhagawati Maniraj; Gropeanu Radu A; Muniz Gemma M Rodriguez; Sundaram Subramanian; Piehler Jacob^*; del Campo Aranzazu

来源：Small, 2013, 9(6): 838-845.

DOI：10.1002/smll.201201901

• protein micropatterns
• laser lithography
• photocleavable peptides
• phototriggers
• histidine tags

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.

• 出版日期2013-3-25