Gold Fluorescent Annexin A5 as a Novel Apoptosis Detection Tool

Kurschus, F. C.; Pal, P. P.; Bäumler, P.; Jenne, D. E.; Wiltschi, B.; Budisa, N.

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Gold Fluorescent Annexin A5 as a Novel Apoptosis Detection Tool

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Kurschus, F. C.
Department: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society;

Pal, P. P.
Max Planck Society;

Bäumler, P.
Max Planck Society;

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Jenne, D. E.
Department: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society;

Wiltschi, B.
Max Planck Society;

Budisa, N.
Max Planck Society;

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Citation

Kurschus, F. C., Pal, P. P., Bäumler, P., Jenne, D. E., Wiltschi, B., & Budisa, N. (2009). Gold Fluorescent Annexin A5 as a Novel Apoptosis Detection Tool. Cytometry Part A, 75A(7), 626-633.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0012-2069-0

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 Ca2+ 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. (C) 2009 International Society for Advancement of Cytometry