Purification of a membrane-derived human erythroid growth factor

The authors have purified erythroid burst-promoting activity (BPA) from human lymphocyte plasma membranes by detergent extraction followed by gel-filtration, ion-exchange, and hydroxylapatite chromatography. BPA is a heat-stable integral membrane glycoprotein of M/sub r/ 28,000 by gel filtration whose activity is eluted from NaDodSO4polyacrylamide gels as a broad band at M/sub r/ 25,000-29,000. The growth stimulator appears to be erythroid-specific, stimulating proliferation of the human erythroid burst-forming unit (BFU-E) by up to 600% of control values when tested in serum-free bone marrow culture. In contrast, it is devoid of granulocytemacrophage colony-stimulating factor activity and has a negligible effect on the formation of human megakaryocyte and mixed hematopoietic colonies. Polyclonal anti-lymphocyte membrane, IgC, which neutralizes BPA expression in culture, completely absorbs BPA from all lymphocyte-derived sources [solubilized lymphocyte plasma membranes, membrane-containing vesicles shed into lymphocyte conditioned medium (LCM) and soluble vesicle-free LCM supernatants], suggesting that soluble and membrane-derived lymphocyte BPA are antigenically related. This membrane glycoprotein may be an important mediator of proximal cellular interactions that are known to promote erythropoiesis in vitro