Investigation of the molecular architecture of spider silk protein assembly using high-resolution fluorescence microscopy
Abstract
Web spiders synthesize tough silk fibers that outperform mechanical properties of man-made hightech threads. Synthetic spider silk has many anticipated applications ranging from helmets over medical implants to wound-healing mesh. These applications are of potential interest to the Army where they foster warfightersÕ protection and performance. Material scientists are trying to reproduce the light-weight, protein-based material in the laboratory but with limited success. Many details of self-assembly of spider silk proteins, so-called spidroins, are unknown. During silk formation spidroins undergo elusive phase and structural transitions in the spinning duct of the animal, which are triggered by a series of mechanical and chemical stimuli. Here, we propose to apply modern solution fluorescence and super-resolution imaging microscopy methods to detect the assembly architecture of spidroins in silk with virtually molecular resolution. Heterologous expression of spidroins in E. coli bacterial cells will be applied to synthesize engineered building blocks. Purified spidroins will be site-specifically modified with differently colored organic fluorescence labels and assembled into silk in-vitro using a microfluidic mixing device that emulates mechanical and chemical stimuli as they happen in a spiderÕs spinning duct. Equilibrium and kinetic solution fluorescence experiments, resonance energy transfer and two-color superresolution fluorescence imaging microscopy will be applied to reveal interaction patterns of spidroins in silk at the molecular level. The response of these interaction patterns to mechanical and chemical stimuli, which are known to modulate mechanical properties of the material, will be studied. The outcome of the project will aid material scientists in tailoring and improving strength and toughness of spider silk protein-based material.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- May 07, 2018
- Source ID
- W911NF1710336
Entities
People
- Hannes Neuweiler
Organizations
- Army Contracting Command
- United States Army
- University of Würzburg