Autores UPV
Gropeanu Mihaela ,
Bhagawati Maniraj ,
Gropeanu Radu A. Gropeanu,
Rodríguez Muñiz Gemma María,
Sundaram Subramanian ,
Piehler Jacob ,
del Campo Aránzazu
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-specifi c 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. Effi cient and stable caging of tris-NTAfunctionalized
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 fi rst
time. In combination with a short peptide tag for a site-specifi c enzymatic coupling
reaction, covalent immobilization of multiple proteins into target micropatterns is
possible under physiological conditions.