Biological characterization of uncleavable plasma membrane-anchored human macrophage colony-stimulating factor

The cell-surface form of human macrophage colony-stimulating factor (CSF-1( 256), M-CSF alpha) is a plasma membrane-anchored transmembrane protein from which the soluble CSF-1 is released by ectodomain proteolytic cleavage. We have previously generated two forms of cell surface CSF-1 which failed to undergo the cleavage by deleting residues 161-165 or residues 159-165 in th e extracellular juxtamembrane region (1). To determine the biologic signifi cance of the ectodomain cleavage, we compared the biosynthesis and biologic activities of uncleavable mutant CSF-1 forms with those of the cleavable w ild-type (WT) CSF-1. We found that the uncleavable CSF-1 forms were able to accumulate on cell surface at about threefold higher level than the cleava ble WT CSF-1 did. We further demonstrated that the uncleavable plasma membr ane-anchored forms of CSF-1 were biologically active in mediating the proli feration of CSF-1-dependent cells as well as the intercellular adhesion bet ween CSF-1 receptor-bearing cells and CSF-1 expressing cells. Furthermore, the adhesive activity of uncleavable CSF-1 forms was about twofold stronger than that of WT CSF-1, which indicated that the ectodomain cleavage system plays an important role in regulating the biologic activities of membrane- anchored CSF-1. (C) 2000 Academic Press.