| Toward understanding the mechanism of chromophore-assisted laser inactivation--evidence for the primary photochemical steps. | |
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MedLine Citation:
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PMID: 15623352 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Chromophore-assisted laser inactivation (CALI) is a light-mediated technique used to selectively inactivate proteins of interest to elucidate their biological function. CALI has potential applications to a wide array of biological questions, and its efficiency allows for high-throughput application. A solid understanding of its underlying photochemical mechanism is still missing. In this study, we address the CALI mechanism using a simplified model system consisting of the enzyme beta-galactosidase as target protein and the common dye fluorescein. We demonstrate that protein photoinactivation is independent from dye photobleaching and provide evidence that the first singlet state of the chromophore is the relevant transient state for the initiation of CALI. Furthermore, the inactivation process was shown to be dependent on oxygen and likely to be based on photooxidation of the target protein via singlet oxygen. The simple model system used in this study may be further applied to identify and optimize other CALI chromophores. |
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Authors:
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Elke Horstkotte; Torsten Schröder; Jens Niewöhner; Erwin Thiel; Daniel G Jay; Stefan W Henning |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Photochemistry and photobiology Volume: 81 ISSN: 0031-8655 ISO Abbreviation: Photochem. Photobiol. Publication Date: 2005 Mar-Apr |
Date Detail:
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Created Date: 2005-06-02 Completed Date: 2005-09-19 Revised Date: 2005-11-17 |
Medline Journal Info:
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Nlm Unique ID: 0376425 Medline TA: Photochem Photobiol Country: United States |
Other Details:
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Languages: eng Pagination: 358-66 Citation Subset: IM |
Affiliation:
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Xerion Pharmaceuticals AG, Munich, Germany. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Absorption Coloring Agents / chemistry Fluoresceins / chemistry, radiation effects Lasers* Light Models, Chemical Oxygen / chemistry Photochemistry Singlet Oxygen / radiation effects Time Factors beta-Galactosidase / antagonists & inhibitors*, chemistry, radiation effects |
| Chemical | |
Reg. No./Substance:
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0/Coloring Agents; 0/Fluoresceins; 17778-80-2/Singlet Oxygen; 7782-44-7/Oxygen; EC 3.2.1.23/beta-Galactosidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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