Labeling of fusion proteins with synthetic fluorophores in live cells.
A general approach for the sequential labeling of fusion proteins of O(6)-alkylguanine-DNA alkyltransferase (AGT) with different fluorophores in mammalian cells is presented. AGT fusion proteins with different localizations in the cell can be labeled specifically with different fluorophores, and the fluorescence labeling can be used for applications such as multicolor analysis of dynamic processes and fluorescence resonance energy transfer measurements. The facile access to a variety of different AGT substrates as well as the specificity of the labeling reaction should make the approach an important tool to study protein function in live cells.
Keppler
Antje
A
Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Pick
Horst
H
Arrivoli
Claudio
C
Vogel
Horst
H
Johnsson
Kai
K
D000818:Animals; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D006224:Cricetinae; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; D019853:O(6)-Methylguanine-DNA Methyltransferase; D002118:Calcium