Abstract
Structural domains of fibronectin (FN) and their ability to associate with cell surface components have been systematically investigated. Plasma FN was cleaved into three structural domains (Mr 150,000-140,000, 40,000, and 32,000) by sequential digestion with trypsin and thermolysin. A single digestion with thermolysin alone generated Mr 150,000-140,000, 40,000, and smaller fragments. With the inclusion of thermolysin, but not with other proteases, one can, with a high yield dissect FN simultaneously into three clearly distinctive functional domains. Of three major fragments only the Mr 40,000 fragment bound to a gelatin column; this fragment contained essentially all of the carbohydrates present in the original FN. In contrast, heparin-binding sites were localized on both the Mr 150,000-140,000 and 32,000 fragments but not on the Mr 40,000 fragment. Only the Mr 150,000-140,000 fragments and intact FN promoted cell spreading, whereas the Mr 40,000 and 32,000 fragments could induce cell attachment but failed to promote cell spreading. These results indicate that FN is composed of (at least) three structural domains that are functionally distinct from each other.