Molecular abnormalities of human plasminogen isolated from synovial fluid of rheumatoid arthritis patients.
Plasminogen (Pg) in the synovial fluid of patients with acute inflammatory disease, including rheumatoid arthritis, osteoarthritis, and gout, was purified by affinity chromatography techniques. The Pg isolated from patients with osteoarthritis and gout has affinities for L-lysine Sepharose and structural properties similar to those of normal plasma Pg. However, Pg from rheumatoid synovial fluids is abnormal in that it does not bind to L-lysine Sepharose. Analyses of rheumatoid synovial fluid Pg purified by immunoaffinity chromatography indicate that the molecule has size and folding properties similar to those of normal plasma Pg. However, we detected abnormalities in the molecule using two different criteria. First, isolated kringles 1-3 fragments are unable to bind L-lysine Sepharose. Second, reactivity toward a monoclonal antibody against a region in kringles 1-3 is decreased. In addition, rheumatoid synovial fluid Pg competes with normal plasma Pg for binding to the receptor on rheumatoid synovial fibroblasts but not on normal synovial fibroblasts. We also found that binding and activation of rheumatoid synovial fluid derived Pg on the surface of rheumatoid synovial fibroblasts elicits a rise in the concentration of cytosolic free Ca2+ similar to that of normal plasma Pg. Our data suggest that the inability of rheumatoid synovial fluid Pg to bind to L-lysine Sepharose is due to minor structural changes in kringle 1-3. These changes do not affect the physiological properties or the binding ability of synovial fluid Pg to cellular receptors on cells obtained from rheumatoid synovial tissue.
Shearin
T V
TV
Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA.
Pizzo
S V
SV
Gonzalez-Gronow
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000378:metabolism*; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006073:Gout / Q000097:blood; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008297:Male; D010003:Osteoarthritis / Q000097:blood; D010958:Plasminogen / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011817:Rabbits; D013582:Synovial Fluid / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010958:Plasminogen; D008239:Lysine; D002118:Calcium