Photocage-Selective Capture and Light-Controlled Release of Target Proteins
Rasa Rakauskaitė,
Giedrė Urbanavičiūtė,
Martynas Simanavičius,
Rita Lasickienė,
Aušra Vaitiekaitė,
Gražina Petraitytė,
Viktoras Masevičius,
Aurelija Žvirblienė,
Saulius Klimašauskas
Affiliations
Rasa Rakauskaitė
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania
Giedrė Urbanavičiūtė
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania
Martynas Simanavičius
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania
Rita Lasickienė
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania
Aušra Vaitiekaitė
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania
Gražina Petraitytė
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania; Institute of Chemistry, Department of Chemistry and Geosciences, Vilnius University, LT-10257 Vilnius, Lithuania
Viktoras Masevičius
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania; Institute of Chemistry, Department of Chemistry and Geosciences, Vilnius University, LT-10257 Vilnius, Lithuania
Aurelija Žvirblienė
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania
Saulius Klimašauskas
Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania; Corresponding author
Summary: Photochemical transformations enable exquisite spatiotemporal control over biochemical processes; however, methods for reliable manipulations of biomolecules tagged with biocompatible photo-sensitive reporters are lacking. Here we created a high-affinity binder specific to a photolytically removable caging group. We utilized chemical modification or genetically encoded incorporation of noncanonical amino acids to produce proteins with photocaged cysteine or selenocysteine residues, which were used for raising a high-affinity monoclonal antibody against a small photoremovable tag, 4,5-dimethoxy-2-nitrobenzyl (DMNB) group. Employing the produced photocage-selective binder, we demonstrate selective detection and immunoprecipitation of a variety of DMNB-caged target proteins in complex biological mixtures. This combined orthogonal strategy permits photocage-selective capture and light-controlled traceless release of target proteins for a myriad of applications in nanoscale assays.
