Structural Characterization of Black Widow Spider Dragline Silk Proteins CRP1 and CRP4
Poster Number
15a
Introduction/Abstract
Spider silk is a high-performance material with outstanding strength and toughness, rivaling mechanical properties of high tensile steel and Kevlar™. A previous study used mass spectrometry to identify a family of low molecular weight cysteine rich proteins (CRPs) in the Western black widow spider (Lactrodectus hesperus) dragline silk. In this study, we characterize the secondary structure as well as the thermal and pH stability of purified recombinant CRP1 and CRP4 from E. coli.
Purpose
The purpose of this study is to further investigate the protein components of black widow dragline silk. The identification of CRP1 and CRP4 secondary structure in addition to the evaluation of thermal and pH stability were performed.
Method
Large scale expression of recombinant CRP1 and CRP4 were purified from E. coli. The purified proteins were subjected to varying temperature and pH conditions followed by secondary structure analysis using circular dichroism. Deconvolution of all circular dichroism data was performed using DichroWeb to determine the secondary structure content of the CRP1 and CRP4.
Results
Analysis of the CRPs revealed that both CRP1 and CRP4 have secondary structures that are largely random-coil. Additionally, over a temperature range of 15oC to 85oC at pH 7, and over at pH range of 5.8 to 7.8 at 25oC, little to no structural changes were observed in CRP1 and CRP4.
Significance
Factors effecting the mechanical properties of spider silk can be assessed through characterization of proteins found in the final dragline silk fibers. This study provides biochemical data that can be incorporated synthetic silk production.
Location
DeRosa University Center
Format
Poster Presentation
Poster Session
Afternoon 1pm-3pm
Structural Characterization of Black Widow Spider Dragline Silk Proteins CRP1 and CRP4
DeRosa University Center
Spider silk is a high-performance material with outstanding strength and toughness, rivaling mechanical properties of high tensile steel and Kevlar™. A previous study used mass spectrometry to identify a family of low molecular weight cysteine rich proteins (CRPs) in the Western black widow spider (Lactrodectus hesperus) dragline silk. In this study, we characterize the secondary structure as well as the thermal and pH stability of purified recombinant CRP1 and CRP4 from E. coli.
