Binding sites and endocytosis of heparin and polylysine are changed when the two molecules are given as a complex to Chinese hamster ovary cells.

This paper characterizes the complex formed in vitro between polylysine and heparin in the presence of heparin excess, and investigates the interaction of this complex with the surface of Chinese hamster ovary cells. It examines the kinetics of surface binding and cellular uptake of the complex and shows that both processes can be distinguished from those of free heparin and free polylysine. The view that these three ligands bind to different surface sites is further supported by the fact that their interaction with cells is influenced differently by cell detachment with trypsin, detachment with EGTA or exposure to acid pH. Membrane transport of the complex is a saturable process suggestive of receptor-mediated endocytosis. It is, however, less effective than would be expected on the basis of the binding kinetics. Only 40% of the complex bound at 0 degrees C is internalized during a 2 h reincubation period at 37 degrees C, suggesting some degree of uncoupling between binding and endocytosis. These data confirm prior results obtained with methotrexate-polylysine conjugates. We had shown that the addition of heparin to a medium containing a methotrexate-polylysine conjugate leads unexpectedly to a marked cellular uptake of drug conjugate, which is capable of killing cells that are otherwise resistant to free methotrexate (Shen, W.-C. and Ryser, H.J.-P. (1981) Proc. Natl. Acad. Sci. USA 78, 7589-7593). The polylysine X heparin complex is therefore of interest as a potential carrier for intracellular drug delivery through endocytosis.