Rat osteoclasts adhere to a wide range of RGD (Arg-Gly-Asp) peptide-containing proteins, including the bone sialoproteins and fibronectin, via a beta 3 integrin.
The ligand binding ability of rat osteoclast adhesion receptors was investigated in an attachment assay using osteoclasts disaggregated from bone. Osteoclasts adhered well to the Arg-Gly-Asp (RGD)-containing proteins osteopontin (bone sialoprotein I) and BSP (bone sialoprotein II), vitronectin, fibrinogen, von Willebrand factor, and fibronectin. Osteoclasts also adhered, but less strongly, to type I collagen. No attachment of osteoclasts was observed to thrombospondin, tenascin, laminin, or a range of non-RGD-containing bone proteins and proteins from other sources. The attachment of osteoclasts to all ligands was abolished in the presence of GRGDSP peptide, indicating the involvement of the RGD cell binding sequence in ligand binding. Attachment of osteoclasts to all substrates, with the exception of type I collagen, was also strongly inhibited by the addition of monoclonal antibody F11 to the beta 3 integrin subunit, indicating that a beta 3 integrin, probably the vitronectin receptor, was involved. Attachment to type I collagen was blocked by EDTA chelation of divalent cations and was not significantly affected by anti-beta 3 or anti-beta 1 antibodies; when taken with the inhibition by RGD peptide, this suggests the involvement of various receptors, possibly including nonintegrin collagen receptors, in the binding of osteoclasts to this protein. These results define the wide range of ligands for extracellular matrix receptors in osteoclasts in vitro. It remains to be established which of these proteins are important in osteoclast adhesion and osteoclastic bone resorption in vivo.
Helfrich
M H
MH
Department of Haematology, St. Bartholomew's Hospital, London, England.
Nesbitt
S A
SA
Dorey
E L
EL
Horton
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000911:Antibodies, Monoclonal; D002448:Cell Adhesion / Q000502:physiology; D003094:Collagen / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry; D039661:Integrin beta3; D016023:Integrins / Q000502:physiology; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis*; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012795:Sialoglycoproteins / Q000737:chemistry
D000906:Antibodies; D000911:Antibodies, Monoclonal; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D039661:Integrin beta3; D016023:Integrins; D009842:Oligopeptides; D011506:Proteins; D012795:Sialoglycoproteins; C047981:arginyl-glycyl-aspartic acid; D003094:Collagen