A calcium-binding, asparagine-linked oligosaccharide is involved in skeleton formation in the sea urchin embryo.
We have previously identified a 130-kD cell surface protein that is involved in calcium uptake and skeleton formation by gastrula stage embryos of the sea urchin Strongylocentrotus purpuratus (Carson et al., 1985. Cell. 41:639-648). A monoclonal antibody designated mAb 1223 specifically recognizes the 130-kD protein and inhibits Ca+2 uptake and growth of the CaCO3 spicules produced by embryonic primary mesenchyme cells cultured in vitro. In this report, we demonstrate that the epitope recognized by mAb 1223 is located on an anionic, asparagine-linked oligosaccharide chain on the 130-kD protein. Combined enzymatic and chemical treatments indicate that the 1223 oligosaccharide contains fucose and sialic acid that is likely to be O-acetylated. Moreover, we show that the oligosaccharide chain containing the 1223 epitope specifically binds divalent cations, including Ca+2. We propose that one function of this negatively charged oligosaccharide moiety on the surfaces of primary mesenchyme cells is to facilitate binding and sequestration of Ca+2 ions from the blastocoelic fluid before internalization and subsequent deposition into the growing CaCO3 skeleton.
Farach-Carson
M C
MC
Department of Biological Chemistry, University of Texas Health Science Center, Houston 77225.
Carson
D D
DD
Collier
J L
JL
Lennarz
W J
WJ
Park
H R
HR
Wright
G C
GC
D000818:Animals; D000911:Antibodies, Monoclonal; D001216:Asparagine; D002135:Calcium-Binding Proteins / Q000032:analysis* / Q000096:biosynthesis; D002478:Cells, Cultured; D004625:Embryo, Nonmammalian / Q000502:physiology; D000939:Epitopes / Q000032:analysis; D005775:Gastrula / Q000502:physiology*; D006020:Glycopeptides / Q000302:isolation & purification; D006023:Glycoproteins / Q000032:analysis* / Q000096:biosynthesis; D006026:Glycoside Hydrolases; D015151:Immunoblotting; D009844:Oligosaccharides / Q000032:analysis*; D010012:Osteogenesis; D012617:Sea Urchins / Q000196:embryology*
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D000939:Epitopes; D006020:Glycopeptides; D006023:Glycoproteins; D009844:Oligosaccharides; D001216:Asparagine; D006026:Glycoside Hydrolases