Expression of platelet glycoprotein Ib by cultured human megakaryocytes: ultrastructural localization and biosynthesis.
Glycoprotein Ib (GPIb), the receptor for von Willebrand factor, is a two-chain member constituent of the platelet/megakaryocytic lineage. Studies on its expression have been hampered by the difficulties in obtaining purified megakaryocytes in a sufficient number. We report a suspension liquid culture procedure that allowed isolation of more than 1 x 10(6) megakaryocytes with a purity ranging from 3% to 88% from the blood of patients with chronic myeloid leukemia, from fetal liver or from normal human bone marrow. GPIb was detected on the plasma membrane of all maturing megakaryocytes and also of promegakaryoblasts devoid of demarcation membranes. GPIb was detected on demarcation membranes of maturing megakaryocytes but was absent from all other organelles, including alpha granules. Biosynthesis of 35S-methionine labeled megakaryocytes showed that GPIb with similar electrophoretic mobility to the platelet molecule was synthesized and that it was also composed of two chains, since its molecular weight shifted in reducing conditions from 170 Kd to 145 Kd. The beta chain remained undetectable after methionine metabolic labeling, but it was immunoprecipitated after 3H-leucine metabolic labeling, confirming that this subunit is devoid of methionine. GPIb was associated with GPIX, as it is in platelets, since anti-GPIb antibodies coprecipitated a 17 Kd polypeptide.
Debili
N
N
INSERM U91, Hôpital Henri Mondor, Créteil, France.
Kieffer
N
N
Nakazawa
M
M
Guichard
J
J
Titeux
M
M
Cramer
E
E
Breton-Gorius
J
J
Vainchenker
W
W
D000328:Adult; D001854:Bone Marrow Cells; D002462:Cell Membrane / Q000032:analysis; D002469:Cell Separation; D002478:Cells, Cultured; D003593:Cytoplasm / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007163:Immunosorbent Techniques; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000097:blood; D008099:Liver / Q000166:cytology / Q000196:embryology; D008533:Megakaryocytes / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008970:Molecular Weight; D010980:Platelet Membrane Glycoproteins / Q000032:analysis* / Q000096:biosynthesis
D010980:Platelet Membrane Glycoproteins