Interaction of plasma fibronectin with gelatin and complement C1q.
A variety of techniques have been used to examine the interaction of human plasma fibronectin (Fn) with complement C1q in comparison to that with gelatin in phosphate buffered saline at pH 7.4. The precipitation of 3H-Fn by polyethylene glycol (PEG) was shifted to much lower concentrations of the polymer by addition of gelatin, and to a lesser extent, by C1q. Precipitation of 3H-Fn in the presence of C1q was close to that of C1q alone under identical conditions suggesting an affinity of Fn for solid phase C1q; a similar interaction was seen with heat-insolubilized C1q. Fibronectin bound tightly to gelatin-Sepharose and C1q-Sepharose and this binding could be inhibited by gelatin but not by C1q. The presence of gelatin retarded the anodal migration of Fn during immunoelectrophoresis under physiological conditions whereas C1q had an effect only at low ionic strength. Exclusion chromatography of Fn, alone and preincubated with gelatin or C1q, was also consistent with the formation of strong complexes with gelatin but not with C1q, whereas similar mixtures of Fn and gelatin exhibited a fast-sedimenting boundary and marked depletion of the 12S Fn peak. Titration of fluorescein-labeled alpha 2 chains of type I collagen with Fn produced an increase in fluorescence polarization which could be reversed by addition of unlabeled alpha 2 chains or gelatin but not by C1q or the pepsin-derived collagen-like domain of C1q. These observations indicate that the fluid-phase interaction of Fn with C1q is much weaker than that with gelatin but that Fn does have appreciable affinity for solid-phase C1q. Such interaction could signify a role for Fn in the clearance of immune complexes from circulation.
D000818:Animals; D011232:Chemical Precipitation; D002845:Chromatography; D002846:Chromatography, Affinity; D003166:Complement Activating Enzymes / Q000378:metabolism*; D015922:Complement C1q; D005353:Fibronectins / Q000097:blood*; D005454:Fluorescence Polarization; D005780:Gelatin / Q000378:metabolism*; D006801:Humans; D007122:Immunoelectrophoresis; D011092:Polyethylene Glycols; D051381:Rats; D014461:Ultracentrifugation
D005353:Fibronectins; D011092:Polyethylene Glycols; D015922:Complement C1q; D005780:Gelatin; D003166:Complement Activating Enzymes