Immunological screening of SPARC/Osteonectin in nonmineralized tissues.
SPARC/Osteonectin is a major bone-related protein that is also present in nonmineralized tissues and in platelets. As compared to bone SPARC/Osteonectin, SPARC/Osteonectin from platelets presents a slightly lower electrophoretic mobility in SDS-PAGE and a 100-fold decreased affinity for a unique monoclonal antibody, Mab2 (Malaval et al. 1991). To check the tissular diversity of SPARC/Osteonectin, protein extracts from bovine bone, nonmineralized tissues, and platelets were screened by immunoblotting and immunoradiometric assay, with Mab2 and three other monoclonal antibodies recognizing distinct epitopes. The SPARC/Osteonectin secreted by a human osteosarcoma cell line (MG63) was also tested. In all the nonmineralized tissues tested (gut, bone marrow, tendon, mesentery, artera, lens, skin, liver, and cornea), SPARC/Osteonectin presents the same immunoreactivity and electrophoretic mobility as in bone. The heavier molecular weight and Mab2-negative form present in platelets seems to be unique to this cell type. Osteosarcoma cell extracts and conditioned media give the same results as bone extracts, indicating that the low molecular weight and Mab2-positive form of SPARC/Osteonectin present in most tissues does not result from proteolytic cleavage in the matrix, but is secreted as such. Bone and platelet SPARC/Osteonectin present different patterns of sensitivity to glycosidases, suggestive of a difference in N-glycosylation. However, these treatments do not affect the decreased affinity of Mab2 for platelet SPARC/Osteonectin, which is not likely to be related to difference in N-glycosylation.
Maillard
C
C
Inserm U234, Hopital Edouard Herriot, Lyon, France.
Malaval
L
L
Delmas
P D
PD
D000818:Animals; D000911:Antibodies, Monoclonal; D001345:Autoradiography; D001792:Blood Platelets / Q000737:chemistry; D001842:Bone and Bones / Q000737:chemistry; D002417:Cattle; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005333:Fetus / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D006882:Hydroxyapatites / Q000378:metabolism; D015151:Immunoblotting; D015592:Immunoradiometric Assay; D008970:Molecular Weight; D009928:Organ Specificity; D015676:Osteonectin / Q000737:chemistry / Q000276:immunology*; D012516:Osteosarcoma / Q000737:chemistry; D011863:Radioimmunoassay; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D006882:Hydroxyapatites; D015676:Osteonectin