Using model substrates to study the dependence of focal adhesion formation on the affinity of integrin-ligand complexes.
The adhesion of mammalian cells is mediated by the binding of cell-surface integrin receptors to peptide ligands from the extracellular matrix and the clustering of these receptors into focal adhesion complexes. This paper examines the effect of one mechanistic variable, ligand affinity, on the assembly of focal adhesions (FAs) in order to gain mechanistic insight into this process. This study uses self-assembled monolayers of alkanethiolates on gold as a substrate to present either a linear or cyclic Arg-Gly-Asp peptide at identical densities. Inhibition assays showed that the immobilized cyclic RGD is a higher affinity ligand than linear RGD. 3T3 Swiss fibroblasts attached to substrates presenting the cyclic peptide at twice the rate they attached to substrates presenting the linear peptide. Quantitation of focal adhesions revealed that cells on cyclic RGD had twice the number of FAs as did cells on linear RGD and that these focal adhesions were on average smaller. These findings show that affinity affects the assembly of integrins into focal adhesions and support a model based on competing rates of nucleation and growth of FAs to explain the change in distribution of FAs with ligand affinity. This study is important because it provides a model system that is well-suited for biophysical studies of integrin-mediated cell adhesion and reveals insight into one mechanism utilized by cells to perceive environmental changes.
Department of Chemistry, Institute of Biophysical Dynamics, The University of Chicago, Chicago, Illinois 60637, USA.
Mihoko
M
Kato
Milan
M
Mrksich
D016475:3T3 Cells; D000818:Animals; D001667:Binding, Competitive; D002448:Cell Adhesion; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D022001:Focal Adhesions / Q000737:chemistry* / Q000502:physiology*; D016023:Integrins / Q000737:chemistry* / Q000502:physiology; D008024:Ligands; D051379:Mice; D008956:Models, Chemical; D009842:Oligopeptides / Q000737:chemistry; D010766:Phosphorylation; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000737:chemistry; D011971:Receptors, Immunologic / Q000737:chemistry; D015398:Signal Transduction; D013379:Substrate Specificity
D016023:Integrins; D008024:Ligands; D009842:Oligopeptides; D011971:Receptors, Immunologic; C047981:arginyl-glycyl-aspartic acid; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496051:Ptk2 protein, mouse