Purification of B16-F1 melanoma autocrine motility factor and its receptor.
作者： Steve Silletti , Ivan R. Nabi , Victor Hogan , Hideomi Watanabe , Avraham Raz
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摘要: Tumor autocrine motility factor (AMF) is a cytokine which stimulates both random and directed cell migration by self-producing cells. AMF has been detected in purified from serum-free conditioned medium of murine B16-F1 melanoma Under nonreducing conditions migrates sodium dodecyl sulfate-polyacrylamide gel electrophoresis as single band M r 55,000, whereas under reducing it polypeptide 64,000. Two-dimensional polyacrylamide the resolved two polypeptides with isoelectric points 6.35 (major) 6.4 (minor). No carbohydrate side chains were AMF. Purified stimulated dose-dependent fashion bound directly protein-protein-binding assay to receptor, surface glycoprotein 78,000 [glycoprotein (gp) 78]. The involvement gp78 AMF-stimulated function was demonstrated assays. These results suggest that natural ligand for gp78-AMF receptor.
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