Complex protein composition of isolated focal adhesions: a two-dimensional gel and database analysis.
Current ideas about the composition of the focal adhesion complexes in cultured cells are based mainly on indirect immunocytochemical data. We here report a two-dimensional (2-D) gel electrophoresis analysis of the focal adhesion associated-structures that remain in the growth substrate after removal of cells by mechanical shearing. Many proteins additional to the known adhesion proteins, and in higher abundance, could be identified. Using selective extraction procedures, employing detergent or gelsolin, these could be classified as either membrane-associated, actin-associated or both. Cross correlation of these polypeptode patterns with a 2-D gel database allowed identification of some proteins, not previously considered as resident of focal adhesions. The data point to a more complex make up of focal adhesions than formerly supposed.
Niederreiter
M
M
Institute of Molecular Biology, Austrian Academy of Sciences, Salzburg.
Gimona
M
M
Streichsbier
F
F
Celis
J E
JE
Small
J V
JV
D016475:3T3 Cells; D000199:Actins / Q000032:analysis; D000818:Animals; D001345:Autoradiography / Q000379:methods; D002448:Cell Adhesion; D002460:Cell Line; D002461:Cell Line, Transformed; D002642:Chick Embryo; D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007457:Iodine Radioisotopes; D051379:Mice; D008954:Models, Biological; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D016596:Vinculin / Q000032:analysis
D000199:Actins; D007457:Iodine Radioisotopes; D011506:Proteins; D016596:Vinculin