A brilliant monomeric red fluorescent protein to visualize cytoskeleton dynamics in Dictyostelium.

Abstract:

:Red fluorescent proteins (RFPs) combined with GFP are attractive probes for double-fluorescence labeling of proteins in live cells. However, the application of these proteins is restrained by stable oligomer formation and by their weak fluorescence in vivo. Previous attempts to eliminate these problems by mutagenesis of RFP from Discosoma (DsRed) resulted in the monomeric mRFP1 and in the tetrameric RedStar RFP, which is distinguished by its enhanced fluorescence in vivo. Based on these mutations, we have generated an enhanced monomeric RFP, mRFPmars, and report its spectral properties. Together with green fluorescent labels, we used mRFPmars to visualize filamentous actin structures and microtubules in Dictyostelium cells. This enhanced RFP proved to be suitable to monitor the dynamics of cytoskeletal proteins in cell motility, mitosis, and endocytosis using dual-wavelength fluorescence microscopy.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Fischer M,Haase I,Simmeth E,Gerisch G,Müller-Taubenberger A

doi

10.1016/j.febslet.2004.09.084

keywords:

subject

Has Abstract

pub_date

2004-11-05 00:00:00

pages

227-32

issue

1-2

eissn

0014-5793

issn

1873-3468

pii

S0014579304012414

journal_volume

577

pub_type

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