Title

Structural Characterization of Black Widow Spider Dragline Silk Proteins CRP1 and CRP4

Poster Number

15a

Lead Author Affiliation

Biological Sciences

Lead Author Status

Masters Student

Second Author Affiliation

Biological Sciences

Third Author Affiliation

Biological Sciences

Fourth Author Affiliation

Biological Sciences

Fourth Author Status

Masters Student

Fifth Author Affiliation

Pharmaceutical and Chemical Sciences

Fifth Author Status

Doctoral Student

Sixth Author Affiliation

Biological Sciences

Additional Authors

Simmone Dyrness - Biological Sciences Dr. Craig Vierra - Faculty Department of Biological Sciences

Introduction

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

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Apr 28th, 1:00 PM Apr 28th, 3:00 PM

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.