Extracellular annexin A5: functions of phosphatidylserine-binding and two-dimensional crystallization.

In normal healthy cells phosphatidylserine is located in the inner leaflet of the plasma membrane. However, on activated platelets, dying cells and under specific circumstances also on various types of viable leukocytes phosphatidylserine is actively externalized to the outer leaflet of the plasma membrane. Annexin A5 has the ability to bind in a calcium-dependent manner to phosphatidylserine and to form a membrane-bound two-dimensional crystal lattice.

In vitro measurement of cell death with the annexin A5 affinity assay.

One of the hallmarks of cell death is the cell surface-expression of phosphatidylserine. Expression of phosphatidylserine at the cell surface can be measured in vitro with the phosphatidylserine-binding protein annexin A5 conjugated to fluorochromes. This measurement can be made by flow cytometry or by confocal scanning-laser microscopy.

Annexin A5 inhibits engulfment through internalization of PS-expressing cell membrane patches.

Apoptosis and subsequent clearance of apoptotic cells are important for the prevention of diseases. Therefore, it is essential to understand the mechanisms underlying the biology of phagocytic clearance of apoptotic cells. The best characterized "eat me" signal on the surface of apoptotic cells is phosphatidylserine (PS).

Cell surface-expressed phosphatidylserine and annexin A5 open a novel portal of cell entry.

Expression of phosphatidylserine (PtdSer) at the cell surface is part of the membrane dynamics of apoptosis. Expressed phosphatidylserine functions as an "eat me" flag toward phagocytes. Here, we report that the expressed phosphatidylserine forms part of a hitherto undescribed pinocytic pathway.

Visualization of cell death in vivo with the annexin A5 imaging protocol.

Annexin A5 binds to phosphatidylserine (PS), which is one of the "eat me" signals at the surface of the apoptotic cell. This property has been the driving force for the research of annexin A5 as a probe to measure apoptosis in vitro and in vivo. A non-invasive imaging protocol using annexin A5 has been developed and applied successfully to measure programmed cell death programmed cell death (PCD) in patients.