Topographic changes of focal adhesion components and modulation of p125FAK activation in stretched human periodontal ligament fibroblasts.
Mechanical stress has been shown in vitro to modulate integrin-beta1-mediated activation of p125FAK/FAK. To test the hypothesis whether this also applies to periodontal ligament fibroblasts (PDLs), we subjected human PDLs to mechanical stretch and analyzed stress-induced changes of p125FAK activation by quantitative immunoprecipitation of p125FAK and changes in the topography of molecules localizing in focal adhesions by indirect immunofluorescence. Generally, all components of focal contacts under study-including detection of phosphotyrosine, i.e., integrin-beta1, p125FAK, and paxillin-revealed a relative co-localization during stretch application. Under stretch, we observed a re-distribution of all components from the cell periphery to the cytoplasm following the main axes. Tyrosine phosphorylation of p125FAK was monitored up to 72 hours under stretch. While the amount of p125FAK remained essentially constant, the activation of p125FAK was clearly modulated. Tyrosine phosphorylation of p125FAK increased from 15 minutes up to 1 hour and declined after stretching periods of 24, 48, and 72 hours. The analysis of our data indicated a stretch-induced redistribution of focal adhesion components and a modulation of p125FAK activation, suggesting alterations in focal adhesions and their associated signal cascade.
Molina
T
T
Department of Orthodontics and Dentofacial Orthopedics, Dental School, University of Heidelberg, Germany.
Kabsch
K
K
Alonso
A
A
Kohl
A
A
Komposch
G
G
Tomakidi
P
P
D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003799:Dental Stress Analysis; D004789:Enzyme Activation; D005347:Fibroblasts / Q000201:enzymology / Q000502:physiology; D019084:Fluorescent Antibody Technique, Indirect; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D022001:Focal Adhesions / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D019012:Integrin beta1 / Q000378:metabolism; D051419:Paxillin; D010513:Periodontal Ligament / Q000166:cytology* / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D015398:Signal Transduction; D013314:Stress, Mechanical
D003598:Cytoskeletal Proteins; D019012:Integrin beta1; C496061:PXN protein, human; D051419:Paxillin; D010750:Phosphoproteins; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human