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An endogenous glycosylphosphatidylinositol-specific phospholipase D
releases basic fibroblast growth factor-heparan sulfate proteoglycan
complexes from human bone marrow cultures
G Brunner, CN Metz, H Nguyen, J Gabrilove, SR Patel, MA Davitz, DB Rifkin and EL Wilson
Department of Cell Biology, New York University Medical Center, NY 10016.
Basic fibroblast growth factor (bFGF) is a hematopoietic cytokine that
stimulates stromal and stem cell growth. It binds to a
glycosylphosphatidylinositol (GPI)-anchored heparan sulfate proteoglycan on
human bone marrow (BM) stromal cells. The bFGF- proteoglycan complex is
biologically active and is released by addition of exogenous
phosphatidylinositol-specific phospholipase C. In this study, we show the
presence of an endogenous GPI-specific phospholipase D (GPI-PLD) that
releases the bFGF-binding heparan sulfate proteoglycan and the variant
surface glycoprotein (a model GPI-anchored protein) from BM cultures. An
involvement of proteases in this process is unlikely, because released
proteoglycan contained the GPI anchor component, ethanol-amine, and
protease inhibitors did not diminish the release. The mechanism of release
is likely to involve a GPI-PLD and not a GPI-specific phospholipase C,
because the release of variant surface glycoprotein did not reveal an
epitope called the cross- reacting determinant that is exposed by
phospholipase C-catalyzed GPI anchor cleavage. In addition, phosphatidic
acid (which is specifically a product of GPI-PLD-catalyzed anchor cleavage)
was generated during the spontaneous release of the GPI-anchored variant
surface glycoprotein. We also detected GPI-PLD-specific enzyme activity and
mRNA in BM cells. Therefore, we conclude that an endogenous GPI-PLD
releases bFGF-heparan sulfate proteoglycan complexes from human BM
cultures. This mechanism of GPI anchor cleavage could be relevant for
mobilizing biologically active bFGF in BM. An endogenous GPI-PLD could also
release other GPI-anchored proteins important for hematopoiesis and other
physiologic processes.
Volume 83,
Issue 8,
pp. 2115-2125,
04/15/1994
Copyright © 1994 by The American Society of Hematology
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