Photocage-Selective Capture and Light-Controlled Release of Target Proteins
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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-12-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004220310300 |
| _version_ | 1818846765364805632 |
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| author | Rasa Rakauskaitė Giedrė Urbanavičiūtė Martynas Simanavičius Rita Lasickienė Aušra Vaitiekaitė Gražina Petraitytė Viktoras Masevičius Aurelija Žvirblienė Saulius Klimašauskas |
| author_facet | Rasa Rakauskaitė Giedrė Urbanavičiūtė Martynas Simanavičius Rita Lasickienė Aušra Vaitiekaitė Gražina Petraitytė Viktoras Masevičius Aurelija Žvirblienė Saulius Klimašauskas |
| author_sort | Rasa Rakauskaitė |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-19T05:50:45Z |
| format | Article |
| id | doaj.art-830c0ab1bd404b8f8512da1988cc6b0a |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-12-19T05:50:45Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-830c0ab1bd404b8f8512da1988cc6b0a2022-12-21T20:33:38ZengElsevieriScience2589-00422020-12-012312101833Photocage-Selective Capture and Light-Controlled Release of Target ProteinsRasa Rakauskaitė0Giedrė Urbanavičiūtė1Martynas Simanavičius2Rita Lasickienė3Aušra Vaitiekaitė4Gražina Petraitytė5Viktoras Masevičius6Aurelija Žvirblienė7Saulius Klimašauskas8Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania; Institute of Chemistry, Department of Chemistry and Geosciences, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania; Institute of Chemistry, Department of Chemistry and Geosciences, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, LithuaniaInstitute of Biotechnology, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2589004220310300BiochemistryBiochemistry MethodsBiotechnology |
| spellingShingle | Rasa Rakauskaitė Giedrė Urbanavičiūtė Martynas Simanavičius Rita Lasickienė Aušra Vaitiekaitė Gražina Petraitytė Viktoras Masevičius Aurelija Žvirblienė Saulius Klimašauskas Photocage-Selective Capture and Light-Controlled Release of Target Proteins iScience Biochemistry Biochemistry Methods Biotechnology |
| title | Photocage-Selective Capture and Light-Controlled Release of Target Proteins |
| title_full | Photocage-Selective Capture and Light-Controlled Release of Target Proteins |
| title_fullStr | Photocage-Selective Capture and Light-Controlled Release of Target Proteins |
| title_full_unstemmed | Photocage-Selective Capture and Light-Controlled Release of Target Proteins |
| title_short | Photocage-Selective Capture and Light-Controlled Release of Target Proteins |
| title_sort | photocage selective capture and light controlled release of target proteins |
| topic | Biochemistry Biochemistry Methods Biotechnology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004220310300 |
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