Characterization of the class III collagen receptor, a phosphorylated, transmembrane glycoprotein expressed in nucleated human cells.
We previously identified a 90-kDa cell surface glycoprotein, termed the class III collagen receptor (CRIII), that bound to collagen in affinity chromatography experiments (Wayner, E. A., and Carter, W. G. (1987) J. Cell Biol. 105, 1873-1884). Here, we utilize monoclonal antibodies to define three domains of the CRIII, hydrophobic transmembrane, phosphorylated cytoplasmic, and glycosylated extracellular. The domain designations are based on the following characteristics. (i) Differential extraction, phase partitioning with Triton X-114, and incorporation into liposomes all indicate that the CRIII is an intrinsic membrane receptor with a hydrophobic domain. After incorporation into liposomes the CRIII binds collagen. (ii) Immunofluorescence microscopy reveals that most nucleated cells express the CRIII and that after extraction with Triton X-100, the Triton-insoluble CRIII distributes in a fibrillar pattern at the cell periphery and in closed loops that partially co-distributed with vimentin. The CRIII contains phosphoserine residues which are located on a cytoplasmic domain that may interact with the cytoskeleton. (iii) The CRIII contains 25% carbohydrate in 8-10 asparagine-linked carbohydrate chains of 2800 daltons each bound to a 65-kDa core peptide in the extracellular domain. Peptide mapping with trypsin defined a glycosylated 27-kDa extracellular fragment and a phosphorylated and glycosylated 35-kDa transmembrane fragment. These data suggest a model for the CRIII that links the cytoskeleton with the extracellular matrix.
Carter
W G
WG
Department of Biochemical Oncology, Fred Hutchinson Cancer Research Center, Seattle, Washington.
Wayner
E A
EA
D000469:Alkaline Phosphatase / Q000378:metabolism; D000581:Amidohydrolases / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D008081:Liposomes; D008168:Lung / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D010449:Peptide Mapping; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011956:Receptors, Cell Surface / Q000378:metabolism*; D038981:Receptors, Collagen
D008081:Liposomes; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; D000469:Alkaline Phosphatase; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase