Gold fluorescent annexin A5 as a novel apoptosis detection tool

✍ Scribed by Florian C. Kurschus; Prajna Paramita Pal; Petra Bäumler; Dieter E. Jenne; Birgit Wiltschi; Nediljko Budisa

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 837 KB
• Volume: 75A
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/cyto.a.20737

✦ Synopsis

Abstract

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. © 2009 International Society for Advancement of Cytometry