Phosphatidylserine as an anchor for plasminogen and its plasminogen receptor, histone H2B, to the macrophage surface.
BACKGROUND
Plasminogen (Plg) binding to cell surface Plg receptors (Plg-Rs) on the surface of macrophages facilitates Plg activation and migration of these cells. Histone H2B (H2B) acts as a Plg-R and its cell surface expression is up-regulated when monocytes are differentiated to macrophages via a pathway dependent on L-type Ca(2+) channels and intracellular Ca(2+).


OBJECTIVE
We sought to investigate the mechanism by which H2B, a protein without a transmembrane domain, is retained on the macrophage surface.


METHODS
THP-1 monocytoid cells were induced to differentiate with interferon gamma + Vitamin D3 or to undergo apoptosis by treatment with camptothecin. Flow cytometry and cell surface biotinylation followed by Western blotting were used to measure the interrelationship between Plg binding, cell surface expression of H2B and outer membrane exposure of phosphatidylserine (PS).


RESULTS
H2B interacted directly with PS via an electrostatic interaction. Anti-PS or PS binding proteins, annexin V and protein S, diminished H2B interaction with PS on the surface of differentiated or apoptotic cells and these same reagents inhibited Plg binding to these cells. L-type Ca(2+) channels played a significant role in PS exposure, H2B surface expression and Plg binding induced either by differentiation or apoptosis.


CONCLUSIONS
These data suggest that H2B tethers to the surface of cells by interacting with PS on differentiated or apoptotic monocytoid cells. L-type Ca(2+) channels regulate PS exposure on the surface of these cells. The exposed PS interacts directly with H2B and hence provides sites for Plg to bind to.
Department of Molecular Cardiology, Cleveland Clinic, Joseph J. Jacobs Center for Thrombosis and Vascular Biology, Cleveland, OH 44195, USA.
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D015153:Blotting, Western; D005434:Flow Cytometry; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism*; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding
D006657:Histones; D010718:Phosphatidylserines; D010958:Plasminogen