Structure and reactivity of adsorbed fibronectin films on mica.
Understanding the interactions of adsorbed fibronectin (Fn) with other biomolecules is important for many biomedical applications. Fn is found in almost all body fluids, in the extracellular matrix, and plays a fundamental role in many biological processes. This study found that the structure (conformation, orientation) and reactivity of Fn adsorbed onto mica is dependent on the Fn surface concentration. Atomic force microscopy and x-ray photoelectron spectroscopy were used to determine the surface coverage of adsorbed Fn from isolated molecules at low surface coverage to full monolayers at high surface coverage. Both methods showed that the thickness of Fn film continued to increase after the mica surface was completely covered, consistent with Fn adsorbed in a more upright conformation at the highest surface-Fn concentrations. Time-of-flight secondary ion mass spectrometry showed that relative intensities of both sulfur-containing (cystine, methionine) and hydrophobic (glycine, leucine/isoleucine) amino acids varied with changing Fn surface coverage, indicating that the conformation of adsorbed Fn depended on surface coverage. Single-molecule force spectroscopy with collagen-related peptides immobilized onto the atomic force microscope tip showed that the specific interaction force between the peptide and Fn increases with increasing Fn surface coverage.
Hull
James R
JR
National ESCA and Surface Analysis Center for Biomedical Problems, and Department of Chemical Engineering, University of Washington, Seattle, Washington 98195, USA.
Tamura
Glen S
GS
Castner
David G
DG
D000327:Adsorption; D000538:Aluminum Silicates / Q000737:chemistry*; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D005353:Fibronectins / Q000737:chemistry* / Q000648:ultrastructure*; D008567:Membranes, Artificial; D011485:Protein Binding; D011487:Protein Conformation; D013499:Surface Properties
D000538:Aluminum Silicates; D020099:Coated Materials, Biocompatible; D005353:Fibronectins; D008567:Membranes, Artificial; C011934:mica