Projekt

Focal Adhesion Kinase (FAK) is an important cytoskeletal protein, and as a component of the focal adhesion complex (FAC) becomes phosphorylated and is involved in cell signaling. Recent observation in our laboratory showed that mechanical stimulation of the integrin receptor of mouse embryonic fibroblasts (MEFs) regulates FAK phosphorylation and downstream signaling events which include regulation of proteins like paxillin, ERK and MLCK. The specific aim of the study is to determine how FAK phosphorylation is regulated by force at the molecular level. Magnetic microbeads coated with the integrin ligand RGD will be added to MEFs and a magnetic twisting and pulling device will be used to apply controlled mechanical stresses directly to cell surface integrins. Cells will then be lyzed and Western Blot analysis will be done to detect phosphorylation at the protein level. Frozen stressed/unstressed cell samples will also be viewed by transmission electron microscopy (TEM). These procedures will provide information on the role of focal adhesion kinase in mechanical, structural and regulatory aspects of the membrane FAC linked cytoskeleton.