Direct visualization of collagen-bound proteoglycans by tapping-mode atomic force microscopy.

Most studies on the interaction of collagen with proteoglycans, two universal components of connective tissues, use technical approaches which substantially modify the shape and size of the proteoglycans themselves. In the present study unfixed, untreated collagen fibrils from rat tail tendon were dehydrated and observed by tapping-mode atomic force microscopy. The surface of collagen fibrils immediately reveals a periodic alternation of gap and overlap zones. A thin, transverse ridge decorates the gap zone, while other filamentous structures run on the fibril surface, either parallel or perpendicular to the fibril axis. These surface structures are much enhanced by Cupromeronic Blue preincubation, while pretreatment with chondroitinase ABC removes them completely, leaving barely detectable transverse ridges. The ridge and filaments are likely to represent, respectively, the core protein and the glycosaminoglycan side chains of proteoglycans, displayed with a far better resolution than with conventional histochemical or immunohistochemical techniques. Our data suggest that proteoglycan molecules are capable of different, multiple interactions with the collagen fibril surface as well as with each other.