The biarsenical dye Lumio exhibits a reduced ability to specifically detect tetracysteine-containing proteins within live cells.
Investigating the localisation of proteins within live cells via fluorescence microscopy typically involves the fusion of the protein of interest to a large fluorescent protein such as green fluorescent protein (GFP). Alternate fluorescent labelling technologies such as the fluorescent biarsenical dye molecules (e.g. FlAsH, ReAsH) are preferable to the use of large fusion proteins in many respects and allow greater flexibility in terms of the location of the labelling site. We assessed the ability of the FlAsH-derived biarsenical dye molecule Lumio to label a range of tetracysteine containing proteins within live cells and report that although in some circumstances Lumio is capable of positively detecting such proteins, the sensitivity and specificity of labelling is significantly reduced, making the Lumio-labelling system unsuitable for the detection of a wide range of protein within live cells.
Hearps
Anna C
AC
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
Pryor
Melinda J
MJ
Kuusisto
Henna V
HV
Rawlinson
Stephen M
SM
Piller
Sabine C
SC
Jans
David A
DA
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001152:Arsenicals; D003545:Cysteine / Q000737:chemistry; D005452:Fluoresceins; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D009942:Organometallic Compounds; D010078:Oxazines; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence