mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures.
A monomeric variant of the red fluorescent protein eqFP611, mRuby, is described. With excitation and emission maxima at 558 nm and 605 nm, respectively, and a large Stokes shift of 47 nm, mRuby appears particularly useful for imaging applications. The protein shows an exceptional resistance to denaturation at pH extremes. Moreover, mRuby is about ten-fold brighter compared to EGFP when being targeted to the endoplasmic reticulum. The engineering process of eqFP611 revealed that the C-terminal tail of the protein acts as a natural peroxisomal targeting signal (PTS). Using an mRuby variant carrying the eqFP611-PTS, we discovered that ordered inheritance of peroxisomes is widespread during mitosis of different mammalian cell types. The ordered partitioning is realized by the formation of peroxisome clusters around the poles of the mitotic spindle and ensures that equal numbers of the organelle are inherited by the daughter cells. The unique spectral properties make mRuby the marker of choice for a multitude of cell biological applications. Moreover, the use of mRuby has allowed novel insights in the biology of organelles responsible for severe human diseases.
Kredel
Simone
S
Institute of General Zoology and Endocrinology, University of Ulm, Ulm, Germany.
Oswald
Franz
F
Nienhaus
Karin
K
Deuschle
Karen
K
Röcker
Carlheinz
C
Wolff
Michael
M
Heilker
Ralf
R
Nienhaus
G Ulrich
GU
Wiedenmann
Jörg
J
D000818:Animals; D002460:Cell Line; D022082:Cellular Structures / Q000378:metabolism*; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism*; D051379:Mice; D020675:Peroxisomes / Q000378:metabolism; D013194:Staining and Labeling; D000097573:Red Fluorescent Protein
D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein