Expression of membrane-bound transferrin-like protein p97 on the cell surface of chondrocytes.
A concanavalin-A-binding protein of 76 kDa was purified from the plasma membrane fraction of rabbit chondrocyte cultures. Amino acid sequencing of the N-terminal region and of tryptic peptides of the protein, in addition to sequencing of its cDNA revealed that this protein is highly similar to the tumour-associated antigen p97. Hence, it was concluded that this protein is the rabbit form of p97. Western blotting, Northern blotting and reverse-transcription PCR analyses indicated that rabbit p97 is expressed at high levels in cartilage and chondrocytes, but is barely detectable in the bone, liver, kidney, small intestine, eye, pancreas, heart, testis, skeletal muscle, spleen and fibroblasts. Immunocytochemical and immunohistochemical analyses demonstrated that p97 is expressed in the plasma membrane of chondrocytes. p97 transcript was detected in all zones of the cartilage but the level was relatively low in the hypertrophic zone. These findings suggest that p97 is involved in maintaining the cell surface characteristics of chondrocytes.
Department of Biochemistry, Hiroshima University School of Dentistry, Japan.
T
T
Kawamoto
H
H
Pan
W
W
Yan
H
H
Ishida
E
E
Usui
R
R
Oda
K
K
Nakamasu
M
M
Noshiro
Y
Y
Kawashima-Ohya
M
M
Fujii
H
H
Shintani
Y
Y
Okada
Y
Y
Kato
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm; D000954:Antigens, Surface / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D002462:Cell Membrane / Q000737:chemistry; D002478:Cells, Cultured; D019902:Chondrocytes / Q000737:chemistry*; D003208:Concanavalin A / Q000378:metabolism; D058950:Melanoma-Specific Antigens; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D010449:Peptide Mapping; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011817:Rabbits; D015183:Restriction Mapping; D013499:Surface Properties
D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D003208:Concanavalin A