Synthesis and application of fluorescent ras proteins for live-cell imaging
| Autor: | Herbert Waldmann, Reinhard Reents, Jürgen Kuhlmann, Melanie Wagner, Stefanie Schlummer |
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| Rok vydání: | 2005 |
| Předmět: |
Fluorophore
Microscopy Confocal Chemistry Organic Chemistry Ras Peptide Molecular Sequence Data Protein engineering Protein tag Biochemistry Fluorescence Fluorescence spectroscopy chemistry.chemical_compound Microscopy Fluorescence Live cell imaging ras Proteins Molecular Medicine Animals Amino Acid Sequence BODIPY Molecular Biology Cells Cultured Fluorescent Dyes |
| Zdroj: | Chembiochem : a European journal of chemical biology. 6(1) |
| ISSN: | 1439-4227 |
| Popis: | Semisynthetic Ras proteins are efficient probes for cell-biology experiments. With a Bodipy FL fluorophore introduced at an appropriate site on the Ras peptide by solid-phase synthesis, the resulting Ras chimera is processed by the cellular machinery and the intracellular localization of the protein can then be visualized by means of confocal laser fluorescence microscopy at relatively low concentrations. The absence of a large N-terminal protein tag overcomes possible interferences in the interaction with cellular partner proteins. The fluorescence emission from Bodipy FL is continuous and disappears only after irreversible bleaching. These characteristics make Ras proteins with nonprotein fluorophores suitable for biophysical analysis. The easy accessibility of the lipopeptide moiety by chemical synthesis opens up numerous options for further biological investigations. |
| Databáze: | OpenAIRE |
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