Determination of activated plasma fibronectin using radioactive labelled collagen I.
The plasma concentration of biological active fibronectin was assayed by a protein binding assay using 125I-collagen I as ligand and heparin as activator. The standard curve is linear for a fibronectin range of 1.1-11 pmol (0.5-5.0 micrograms) and the coefficient of variation was less than 10%. The active or activable fibronectin was compared to the immunoreactive fibronectin in plasma from patients with various bacterial diseases. Similar concentrations were detected by the two assays suggesting that all the circulating fibronectin was functionally active. The assay was also applied to determine the structure-function relationship of heparin and heparansulphate in activation of fibronectin. Low-sulphated heparansulphate from umbilical cords and heparin-activated fibronectin but the effect was uncorrelated to anticoagulation activity. Only a small fraction of the heparin was actually capable of activating fibronectin. It is concluded that the assay is very convenient to detect biological active fibronectin and to elucidate the structure-function relationship of heparin and heparansulphate in activating fibronectin.
D000328:Adult; D001424:Bacterial Infections / Q000097:blood / Q000276:immunology; D003094:Collagen / Q000378:metabolism*; D005260:Female; D005353:Fibronectins / Q000097:blood* / Q000276:immunology; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D008297:Male; D008875:Middle Aged; D011869:Radioligand Assay / Q000379:methods*; D013329:Structure-Activity Relationship
D003094:Collagen; D005353:Fibronectins; D006493:Heparin; D006497:Heparan Sulfate; D007457:Iodine Radioisotopes