PADGEM (GMP140) is a component of Weibel-Palade bodies of human endothelial cells.
PADGEM protein (PADGEM), also known as GMP140, is a platelet alpha-granule membrane protein that is translocated to the external membrane after platelet activation. Although the biosynthesis of this protein was originally thought to be confined to megakaryocytes, the synthesis of PADGEM in endothelial cells was recently demonstrated (McEver et al: Blood 70:1974a, 1987). We now describe the subcellular localization of this protein in endothelial cells. Immunofluorescence staining of permeabilized human umbilical vein endothelial cells with KC4, a well characterized monoclonal antibody to PADGEM, showed positively stained elongated structures similar in distribution and shape to Weibel-Palade bodies. Their identity as Weibel-Palade bodies was confirmed by double label immunofluorescence using KC4 and a polyclonal antiserum to von Willebrand factor (vWf), a protein known to be specifically stored in these organelles. All Weibel-Palade bodies were found to contain PADGEM. In contrast to strong perinuclear staining produced with anti-vWf antibodies, no significant perinuclear staining was obtained with KC4, indicating that relatively little PADGEM is present in the endoplasmic reticulum and in the Golgi apparatus. In endothelial cells treated with secretagogues that stimulate vWf release the elongated structures positive for PADGEM disappeared, further identifying these structures as Weibel-Palade bodies. This observation extends the parallels between Weibel-Palade bodies and alpha-granules and suggests a possible functional association between vWf and PADGEM.
Bonfanti
R
R
Center for Hemostasis and Thrombosis Research, New England Medical Center, Boston, MA 02111.
Furie
B C
BC
Furie
B
B
Wagner
D D
DD
D000001:Calcimycin; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000502:physiology; D004730:Endothelium, Vascular / Q000032:analysis* / Q000502:physiology; D006801:Humans; D015388:Organelles / Q000032:analysis* / Q000502:physiology; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000032:analysis*; D014841:von Willebrand Factor / Q000032:analysis
D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D014841:von Willebrand Factor; D000001:Calcimycin