Gold fluorescent annexin A5 as a novel apoptosis detection tool.
We describe a golden fluorescent apoptosis detection tool, which we generated by a fusion of golden fluorescent protein (GdFP) with human annexin A5 (anxA5). GdFP was obtained by replacement of tryptophan at position 66 with 4-aminotryptophan in the chromophore of enhanced cyan fluorescent protein. The GdFP-anxA5 construct combines highly desirable features originating from both fusion partners. These include (i) strong binding to membrane phosphatidylserine patches of apoptotic cells in the presence of Ca(2+) which is brought about by anxA5, (ii) the stable and homogeneous monomeric state, (iii) as well as the red-shifted fluorescence maximum at 574 nm originating from GdFP. We found that GdFP-anxA5 is equally well applicable for apoptosis studies as a routinely used fluorescein 5'-isothiocyanate-annexin A5 conjugate. Golden fluorescent annexin A5 represents a new, stable, and homogeneous red-shifted optical probe for the efficient detection of apoptosis by fluorescence microscopy or by flow cytometry.
Kurschus
Florian C
FC
Max Planck Institute of Neurobiology, Department of Neuroimmunology, Martinsried, Germany. kurschus@uni-mainz.de
Pal
Prajna Paramita
PP
Bäumler
Petra
P
Jenne
Dieter E
DE
Wiltschi
Birgit
B
Budisa
Nediljko
N
D017304:Annexin A5 / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D006046:Gold; D006801:Humans; D018613:Microscopy, Confocal; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism
D017304:Annexin A5; D015415:Biomarkers; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; D006046:Gold