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Fluoroaluminate Stimulates Phosphorylation of p130 Cas and Fak and Increases Attachment and Spreading of Preosteoblastic MC3T3-E1 Cells

Fluoroaluminate Stimulates Phosphorylation of p130 Cas and Fak and Increases Attachment and Spreading of Preosteoblastic MC3T3-E1 Cells

F Freitas, M Jeschke, I Majstorovic, D R Mueller, P Schindler, H Voshol, J Van Oostrum, M Susa

Bone. 2002 Jan;30(1):99-108.

PMID: 11792571

Abstract:

Fluoroaluminate is a G-protein activator, it stimulates osteoblastic cells in culture, and is a bone-forming agent in vivo. To elucidate the mechanisms of G-protein-mediated action of fluoroaluminate in osteoblasts, we studied protein tyrosine phosphorylation in the preosteoblastic cell line MC3T3-E1. Fluoroaluminate, lysophosphatidic acid (LPA; an agonist for G-protein-coupled receptor), or adhesion to type I collagen all stimulated phosphorylation of a similar set of proteins, including p130, p120, p110 (previously identified as proline-rich tyrosine kinase 2, Pyk2), and p70. The phosphorylation of these proteins was sensitive to an Src inhibitor, but not to a Gi-protein inactivator, pertussis toxin. By purification/mass spectrometry and by immunodepletion, p130 protein was identified as p130 Cas (Crk-associated protein), a Src substrate and a protein involved in signaling by cell-adhesion receptors, integrins. Phosphorylation of immunoprecipitated p130 Cas increased upon stimulation with fluoroaluminate and with agonists of G-protein-coupled receptors, but not with growth factors. By immunodepletion, the p120 protein was identified as focal adhesion kinase, Fak. The addition of fluoroaluminate during cell attachment to type I collagen further stimulated phosphorylation of p130 Cas and of Fak. Simultaneously, fluoroaluminate increased the number of attached MC3T3-E1 cells and their spreading. These novel aspects of fluoroaluminate action in cell culture may be important for the bone-forming action of fluoroaluminate in vivo.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP179248590-B	Src Kinase Inhibitor I - CAS 179248-59-0		179248-59-0	Price
AP4506665-B	Focal Adhesion Kinase Inhibitor I - CAS 4506-66-5		4506-66-5	Price

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