Correlating mechanical properties with aggregation processes in electrochemically fabricated collagen membranes.

We show that mechanical stiffness is a useful metric for characterizing complex collagen assemblies, providing insight about aggregation products and pathways in collagen-based materials. This study focuses on mechanically robust collagenous membranes produced by an electrochemical synthesis process. Changing the duration of the applied electric field, or adjusting the electrolyte composition (by adding Ca(2+), K(+), or Na(+) or by changing pH), produces membranes with a range of Young's moduli as determined from force-displacement measurements with an atomic force microscope. The structural organization, characterized by UV-visible spectroscopy, Raman spectroscopy, optical microscopy, and atomic force microscopy, correlates with the mechanical stiffness. These data provide insights into the relative importance of different aggregation pathways enabled by our multiparameter electrochemically induced collagen assembly process.

Biochemical analysis of the interaction of calcium with toposome: a major protein component of the sea urchin egg and embryo.

We have investigated the biochemical and functional properties of toposome, a major protein component of sea urchin eggs and embryos. Atomic force microscopy was utilized to demonstrate that a Ca(2+)-driven change in secondary structure facilitated toposome binding to a lipid bilayer. Thermal denaturation studies showed that toposome was dependent upon calcium in a manner paralleling the effect of this cation on secondary and tertiary structure. The calcium-induced, secondary, and tertiary structural changes had no effect on the chymotryptic cleavage pattern. However, the digestion pattern of toposome bound to phosphatidyl serine liposomes did vary as a function of calcium concentration. We also investigated the interaction of this protein with various metal ions. Calcium, Mg(2+), Ba(2+), Cd(2+), Mn(2+), and Fe(3+) all bound to toposome. In addition, Cd(2+) and Mn(2+) displaced Ca(2+), prebound to toposome, while Mg(2+), Ba(2+), and Fe(3+) had no effect. Collectively, these results further enhance our understanding of the role of Ca(2+) in modulating the biological activity of toposome.