Purification and characterization of an abundant cytosolic protein from human neutrophils that promotes Ca2(+)-dependent aggregation of isolated specific granules.
Intracellular ionized calcium has been strongly implicated in mediating several responses of human neutrophils to stimulation. However, proteins that serve as effectors of these responses have not been well characterized. To identify proteins that might serve as mediators of the effects of Ca2+ in human neutrophils, we isolated proteins that bind to membrane phospholipids in a Ca2(+)-dependent manner. The most abundant of these, a protein of 33 kD, was readily purified to homogeneity, and was found to bind to phosphatidylserine vesicles in the presence of 2 microM ionized Ca2+. In addition, this purified protein promoted Ca2(+)-dependent aggregation of isolated specific granules from human neutrophils, indicating that it might mediate membrane-membrane contact during processes such as phagosome-lysosome fusion or degranulation. This protein was localized to the cytoplasm of unstimulated neutrophils and found to account for approximately 1% of the cytosol protein. Amino acid sequence of several peptides derived from the purified protein revealed that it is identical to lipocortin III, a recently described member of the annexin family that is scarce in other cells and tissues. The abundance of this protein, together with its Ca2(+)-dependent membrane effects, suggest that it mediates membrane-localized events in stimulated neutrophils, such as phagosome-lysosome fusion or degranulation.
Ernst
J D
JD
Department of Medicine, Rosalind Russell Arthritis Research Laboratory, San Francisco, California.
Hoye
E
E
Blackwood
R A
RA
Jaye
D
D
D000595:Amino Acid Sequence; D017302:Annexins; D001798:Blood Proteins / Q000302:isolation & purification*; D002118:Calcium / Q000502:physiology*; D002135:Calcium-Binding Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D002449:Cell Aggregation / Q000187:drug effects; D003594:Cytoplasmic Granules / Q000187:drug effects*; D003600:Cytosol / Q000032:analysis; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000032:analysis*; D010743:Phospholipids / Q000378:metabolism
D017302:Annexins; D001798:Blood Proteins; D002135:Calcium-Binding Proteins; D010743:Phospholipids; D002118:Calcium