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Synthesis and Application of Fluorescent Ras Proteins for Live‐Cell Imaging
Author(s) -
Reents Reinhard,
Wagner Melanie,
Schlummer Stefanie,
Kuhlmann Jürgen,
Waldmann Herbert
Publication year - 2005
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200400233
Subject(s) - bodipy , fluorophore , fluorescence , chemical biology , biophysics , confocal microscopy , fluorescence microscope , chemistry , protein tag , chimera (genetics) , moiety , peptide , green fluorescent protein , biochemistry , microbiology and biotechnology , fusion protein , biology , stereochemistry , recombinant dna , physics , quantum mechanics , gene
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.