Differential phosphorylation of paxillin in response to surface-bound serum proteins during early osteoblast adhesion.
An early signaling event during the adhesion and spreading of cells is integrin-mediated tyrosine phosphorylation of the cytoskeletal adaptor protein paxillin and the non-receptor tyrosine kinase pp125(FAK) at focal contacts. To determine the influence of surface-charge and -adsorbed adhesion proteins on this signaling pathway, paxillin phosphorylation was examined during attachment of MC3T3-E1 osteoblast-like cell onto charged and uncharged polystyrene, and on adsorbed layers of serum proteins, fibronectin (Fn), vitronectin (Vn), a mixture of Fn and Vn, and albumin. Paxillin phosphorylation was induced 2.4-fold (P < 0.05) on charged vs uncharged polystyrene only in the presence of serum proteins. Activation of paxillin via Fn or Vn alone, or in combination, resulted in significantly lower phosphorylation signals compared to whole serum (41 +/- 6.9%, P < 0.05, 45 +/- 5.9%, P < 0.05, and 76 +/- 9.8%, P < 0.075, respectively). Confocal laser microscopy confirmed increased co-localization of phosphotyrosine and paxillin at protruding lamellopodia of spreading osteoblasts on charged vs uncharged serum-pretreated polystyrene. Taken together, these data suggest that subtle differences in surface characteristics mediate effects on adhering cells via adsorbed serum proteins involving the cytoskeletal adaptor protein paxillin.
Sommerfeldt
D W
DW
Department of Orthopaedics and Biomedical Engineering, State University of New York at Stony Brook, Stony Brook, NY 11794, USA.
McLeod
K J
KJ
Rubin
C T
CT
Hadjiargyrou
M
M
D016475:3T3 Cells; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001798:Blood Proteins / Q000502:physiology*; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002462:Cell Membrane / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000502:physiology; D051379:Mice; D018613:Microscopy, Confocal; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology*; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011137:Polystyrenes; D012709:Serum Albumin / Q000502:physiology; D019096:Vitronectin / Q000502:physiology
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; D005353:Fibronectins; D051419:Paxillin; D010750:Phosphoproteins; D011137:Polystyrenes; C496062:Pxn protein, mouse; D012709:Serum Albumin; D019096:Vitronectin; D019000:Phosphotyrosine