title: Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells.
abstract: The Ca(2+)-dependent binding of Annexin V to phosphatidylserine on cell surfaces is a reliable marker for apoptosis that is widely used in flow cytometry based apoptosis assays. In this paper, we report a new class of Annexin V-based probes for apoptosis. Luciferase from Renilla reniformis (RLuc) was linked to Annexin V and expressed successfully in a soluble form in Escherichia coli BL21 (DE3). The new probe, Rluc/Annexin V, was purified and functionally assayed for detection of apoptosis in actinomycin D-induced apoptotic Jurkat cells. Moreover, the spontaneous apoptosis in neutrophils was shown using the new probe. The results indicate that Rluc/Annexin V can bind to the apoptotic cells, and the signal of Renilla luciferase can be detected by luminometric measurements. The availability of Rluc/Annexin V may be of potential commercial interest for improving current apoptosis assays.
authors.affiliation: Department of Recombinant Technology Research, Nanobiotechnology Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran.
authors.forename: Mahboobeh
authors.initials: M
authors.lastname: Nazari
authors.forename: Rahman
authors.initials: R
authors.lastname: Emamzadeh
authors.forename: Saman
authors.initials: S
authors.lastname: Hosseinkhani
authors.forename: Luca
authors.initials: L
authors.lastname: Cevenini
authors.forename: Elisa
authors.initials: E
authors.lastname: Michelini
authors.forename: Aldo
authors.initials: A
authors.lastname: Roda
mesh_terms: D017304:Annexin A5 / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D006801:Humans; D019169:Jurkat Cells; D049410:Luciferases, Renilla / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015335:Molecular Probes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D017304:Annexin A5; D000970:Antineoplastic Agents; D015335:Molecular Probes; D011993:Recombinant Fusion Proteins; D049410:Luciferases, Renilla