Three-dimensional contact with type I collagen mediates tyrosine phosphorylation in primary human fibroblasts.
By applying Western blot analysis using anti-phosphotyrosine antibodies, primary human dermal fibroblasts were examined after having been cultured on type I collagen-coated surfaces or in free-floating type I collagen gels. In both systems cells showed enhanced tyrosine phosphorylation of a M(r) 120,000 protein (pp120) and of a M(r) 42,000 protein (pp42). Phosphorylation was apparent 6 h at the latest after initiation of the culture and was only slightly induced on polylysine or on plastic. In contrast to pp42, pp120 was rapidly dephosphorylated in cells suspended by trypsinization or released from collagen gels by collagenase treatment, but regained phosphorylation in cells cultured in/on type I collagen. Two human sarcoma cell lines (HT-1080 and RD) exhibited identical tyrosine phosphorylation of pp120 but not of pp42. pp120 is identical with pp125FAK, a novel tyrosine kinase localized in focal adhesions, as proved by immunological cross-reactivity with anti-pp125FAK antibodies. Our results suggest that tyrosine phosphorylation is involved in signal transduction triggered by two- and three-dimensional type I collagen-fibroblast contact.
Roeckel
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D
Department of Dermatology, University of Cologne, Germany.
Krieg
T
T
D000328:Adult; D015153:Blotting, Western; D002448:Cell Adhesion; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis / Q000378:metabolism* / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism*; D005782:Gels; D006801:Humans; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D010766:Phosphorylation; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D012509:Sarcoma / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction / Q000502:physiology; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism*
D005782:Gels; D014443:Tyrosine; D003094:Collagen; D011505:Protein-Tyrosine Kinases