Absence of cell-surface annexin V is accompanied by defective collagen matrix binding in the Swarm rat chondrosarcoma.
Annexin V has been characterized as a major collagen type II binding cell-surface component of normal chondrocytes and is also called anchorin CII in chondrogenic populations. Herein we present evidence that in vitro cultured Swarm rat chondrosarcoma cells are not capable of binding collagen type II in significant quantities to their surfaces, as compared to normal rat chondrocytes. This finding coincides with a deficiency of annexin V on the surface of these cells. A small quantity of an intracellular polypeptide could be detected which is immunologically cross-reactive with annexin V but displayed a mobility in SDS-PAGE of less than 34 kD compared to the M(r) 36 kD of intact rat annexin V. By immunohistochemistry the protein could be localized in the cytoplasm of in vitro and in vivo grown tumor cells. By reverse transcription-polymerase chain reaction and Northern blot analysis, a regular-sized mRNA for annexin V could be detected in the chondrosarcoma cells that is expressed in only slightly lower quantities than in normal chondrocytes. Taken together, the data suggest a modified processing or turnover for annexin V in the chondrosarcoma excluding it from being a functionally active collagen type II binding protein. The findings support the hypothesis of cell-surface annexin V as a key component for the formation of the pericellular matrix of chondrocytes.
King
K B
KB
Department of Biochemistry, Rush Medical College, Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612, USA.
Chubinskaya
S
S
Reid
D L
DL
Madsen
L H
LH
Mollenhauer
J
J
D000595:Amino Acid Sequence; D000818:Animals; D017304:Annexin A5 / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D002645:Chickens; D002813:Chondrosarcoma / Q000737:chemistry / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism*; D003593:Cytoplasm / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured
D017304:Annexin A5; D010446:Peptide Fragments; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003094:Collagen