Adsorption characteristics of plasma fibronectin in relationship to biological activity.
We studied the adsorption properties of plasma fibronectin (pFN) on wettable tissue culture (TC) dishes and nonwettable bacteriological (BAC-T) dishes in relationship to its biological activity of promoting fibroblast spreading. The binding of pFN to the dish surfaces was found to be very tight and partially resistant to treatments with 1M NaOH, 2% SDS, 8M urea, or 6M guanidine HCl. Only the combination of trypsin (1 mg/ml) followed by 1M NaOH resulted in complete recovery of surface bound material. The amount of surface bound pFN did not directly correlate with its activity when comparing TC and BAC-T dishes. At low concentrations, more pFN adsorbed onto the surfaces of BAC-T dishes than TC dishes, but the adsorbed material was biologically less active on BAC-T dishes. At high concentrations, pFN adsorbed similarly on both dish surfaces reaching a maximum level of 320 ng/cm2 or 4.4 x 10(11) molecules/cm2. The possibility was considered that pFN unfolded into an inactive conformation when adsorbed on BAC-T dishes at a low concentration but that at high concentrations, molecular packing requirements prevented unfolding. Evidence in favor of this hypothesis was the observation that addition of 50 micrograms/ml of serum albumin simultaneously with low pFN concentrations resulted in adsorption of pFN on BAC-T dishes in an active conformation even though the amount of adsorbed pFN decreased. Competition experiments between pFN and other proteins were carried out, and it was found that pFN has a much higher affinity for BAC-T or TC dish surfaces than albumin or fibrinogen. It was also found that mild heat denaturation of albumin increased its affinity for the surface by an order of magnitude.
D000327:Adsorption; D004864:Equipment and Supplies; D005340:Fibrinogen / Q000378:metabolism; D005353:Fibronectins / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration
D005353:Fibronectins; D005340:Fibrinogen