Conformational dynamics of amyloid beta-protein assembly probed using intrinsic fluorescence.

Formation of toxic oligomeric and fibrillar structures by the amyloid beta-protein (Abeta) is linked to Alzheimer's disease (AD). To facilitate the targeting and design of assembly inhibitors, intrinsic fluorescence was used to probe assembly-dependent changes in Abeta conformation. To do so, Tyr was substituted in Abeta40 or Abeta42 at position 1, 10 (wild type), 20, 30, 40, or 42. Fluorescence then was monitored periodically during peptide monomer folding and assembly. Electron microscopy revealed that all peptides assembled readily into amyloid fibrils. Conformational differences between Abeta40 and Abeta42 were observed in the central hydrophobic cluster (CHC) region, Leu17-Ala21. Tyr20 was partially quenched in unassembled Abeta40 but displayed a significant and rapid increase in intensity coincident with the maturation of an oligomeric, alpha-helix-containing intermediate into amyloid fibrils. This process was not observed during Abeta42 assembly, during which small decreases in fluorescence intensity were observed in the CHC. These data suggest that the structure of the CHC in Abeta42 is relatively constant within unassembled peptide and during the self-association process. Solvent accessibility of the Tyr ring was studied using a mixed solvent (dimethyl sulfoxide/water) system. [Tyr40]Abeta40, [Tyr30]Abeta42, and [Tyr42]Abeta42 all were relatively shielded from solvent. Analysis of the assembly dependence of the site-specific intrinsic fluorescence data suggests that the CHC is particularly important in controlling Abeta40 assembly, whereas the C-terminus plays the more significant role in Abeta42 assembly

Ultra-sensitive Infrared Spectroscopy of Proteins with Collective Excitations of Nanoplasmonic Arrays

Short interaction lengths limit the application of infrared absorption spectroscopy to the study monolayer thickness films. We employ periodic infrared antenna arrays to obtain 10(4)-10(5) enhancement of protein absorption signals corresponding to zepto-mole sensitivity

Bioactive “self-sensing” optical systems

Free-standing silk films are useful materials to manufacture nanopatterned optical elements and to immobilize bio-dopants such as enzymes while maintaining their biological activity. These traits were combined by incorporating hemoglobin into free-standing silk diffraction gratings to fabricate chemically responsive optofluidic devices responsive to ambient gas conditions, constituting a simple oxygen sensor. This type of self-analyzing optical system is enabled by the unique ability to reproduce high-fidelity optical structures in silk while maintaining the activity of entrapped proteins such as hemoglobin. These bioactive optical devices offer a direct readout capability, adding utility into the bioresponsive material arena