Analysis of the proteins, glycoproteins and glycosaminoglycans of fibroblast adhesions to substratum.
The focal adhesion preparations which remain attached to a glass substratum when fibroblast bodies are removed by a gentle stream of buffer have been analysed by gel electrophoresis coupled with other selective methods of analysis. The results are consistent with the presence of three classes of macromolecular components. (i) Muscle and associated proteins amongst which actin was abundant with significant amounts of tropomyosin, some myosin and traces of alpha-actinin. Some vimentin was present but no vinculin. We detected a major new protein component, as yet unidentified, with a molecular weight in the region of 50000-55000 which is not desmin or tubulin and could have an important function at the focal adhesion. (ii) Glycoproteins which are a specialised subset of those in the whole plasma membrane and included a family which bind ricin and therefore contain beta-galactose end groups, together with a series having carbohydrate chains which bound neither ricin nor concanavalin A. The relative proportion of ricin-binding glycoproteins compared to concanavalin A-binding glycoproteins was higher than in whole plasma membranes. (iii) Glycosaminoglycans, with hyaluronate identified as the major component by column chromatography and its susceptibility to Streptomyces hyaluronidase.
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D005347:Fibroblasts / Q000502:physiology; D006023:Glycoproteins / Q000302:isolation & purification* / Q000502:physiology; D006025:Glycosaminoglycans / Q000302:isolation & purification* / Q000502:physiology; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000502:physiology; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification; D051381:Rats; D011975:Receptors, Mitogen / Q000302:isolation & purification; D012879:Skin Physiological Phenomena
D006023:Glycoproteins; D006025:Glycosaminoglycans; D008565:Membrane Proteins; D009124:Muscle Proteins; D011975:Receptors, Mitogen