Stability of fibronectin biological activity following chemical modification.
Fibronectin isolated from human plasma functions in vitro as a mediator of adhesion and spreading of trypsinized fibroblasts on native or denatured collagen. As a means of elucidating structural characteristics which might contribute to fibronectin's biological activity, we have modified and digested the protein with several chemicals. Following various treatments, the protein was utilized to mediate cell adhesion and spreading on collagen to determine which alteration disrupted its activity. Fibronectin remained functionally intact after partial or complete reduction and alkylation, oxidation of 59% of the carbohydrates with sodium periodate, citraconylation, carbodiimide-catalyzed amide formation, and oxidation of 35.2 residues of tryptophan/molecule with N-bromosuccinimide. Dinitrofluorobenzene treatment, which phenylated ten residues/molecule of fibronectin, successfully inactivated fibronectin's in vitro biological function. Effective modification of the protein was determined by appropriate analytical procedures. Since fibronectin retained its biological function after several treatments that presumably affected its molecular conformation, we concluded that its secondary or tertiary structure appears not to be essential for its in vitro activity, or alternatively that the protein possesses a biologically active domain relatively resistant to chemical modification.
D001974:Bromosuccinimide / Q000494:pharmacology; D002234:Carbodiimides / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D004139:Dinitrofluorobenzene / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology; D004355:Drug Stability; D005029:Ethylenediamines / Q000494:pharmacology; D005353:Fibronectins / Q000097:blood / Q000494:pharmacology; D006801:Humans; D007122:Immunoelectrophoresis; D007460:Iodoacetamide / Q000494:pharmacology; D008970:Molecular Weight; D009578:Nitrobenzenes / Q000494:pharmacology*; D011485:Protein Binding
D002234:Carbodiimides; D005029:Ethylenediamines; D005353:Fibronectins; D009578:Nitrobenzenes; D004139:Dinitrofluorobenzene; D001974:Bromosuccinimide; D004229:Dithiothreitol; D007460:Iodoacetamide