Identification of cytoskeletal components involved in attachment of L929 cells and macrophages to polystyrene.
We have previously shown that lactoperoxidase (LPO) covalently coupled to polystyrene tissue culture flasks can be used to radioiodinate monolayer cell proteins that come into intimate contact with the LPO-polystyrene surface. These studies have now been extended to include a detailed examination of the class of iodinated polypeptides migrating with apparent molecular weights of 50,000 and 55,000 in SDS polyacrylamide gels. Whereas in cultured L929 cells the 55,000 band is predominantly iodinated, in thioglycollate-activated murine peritoneal macrophages the 55,000 and 50,000 bands are of equal intensity. It is possible that the marked degree of exposure of the 50,000 mol wt polypeptide to immobilized LPO is related to the unique strength of macrophages attachment. After labeling of both L929 cells and macrophages with immobilized LPO, all polypeptides in this molecular weight region were subjected to peptide mapping by simultaneous limited proteolysis and electrophoresis in a second SDS polyacrylamide slab gel. The results clearly show that the two major polypeptides in this region are identical within the limits of resolution of this technique. The 55,000 mol wt polypeptide can also be identified in Triton X-100 cytoskeletons from L929 cells after labeling with soluble LPO either before or after detergent lysis. We conclude that this cell surface polypeptide is in continuity with the cytoskeleton and is preferentially exposed to the substratum during attachment to polystyrene.
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D003599:Cytoskeleton / Q000502:physiology*; D007784:Lactoperoxidase; D008264:Macrophages / Q000502:physiology*; D008565:Membrane Proteins / Q000032:analysis / Q000502:physiology*; D051379:Mice; D008970:Molecular Weight; D011137:Polystyrenes
D008565:Membrane Proteins; D011137:Polystyrenes; D007784:Lactoperoxidase