Title

In-Depth Proteome Analysis of Black Widow Dragline Spider Silk

Poster Number

17

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Spider silk in Lactrodectus hesperus (black widow spider) species is renowned for its tensile strength, toughness and elasticity. These properties allow spider silk to outperform some of the best man-made materials such as Kevlar. Despite 6-7 different fiber types, our lab focuses mainly on Major Ampullate (MA) silk. In particular, we are involved in studying the structural proteins (spidroins), which are the molecules that comprise a vast amount of the silk. Previous proteome analysis of dragline silk has revealed approximately 48 proteins - most are uncharacterized. We hypothesize that our proteome analysis of dragline silk will provide a more comprehensive list of proteins, further revealing unknown proteins that are constituents of dragline silk. In order to discover new proteins, we dissolved dragline silk from black widow spiders with urea to denature the proteins. Proteins were further unfolded using reducing agent, followed by alkylation of cysteine sidechain groups. Following these steps, the proteins were digested with trypsin and the peptides separated using nano high performance liquid chromatography. Peptides were subject to MS and MS/MS analysis using an Orbitrap Fusion mass spectrometer. Spectra were analyzed using Proteome Discover 2.1 software. From our experiment we have generated a new list of proteins that more accurately depicts proteins within dragline silk.

Location

DeRosa University Center, Ballroom

Start Date

30-4-2016 10:00 AM

End Date

30-4-2016 12:00 PM

This document is currently not available here.

Share

COinS
 
Apr 30th, 10:00 AM Apr 30th, 12:00 PM

In-Depth Proteome Analysis of Black Widow Dragline Spider Silk

DeRosa University Center, Ballroom

Spider silk in Lactrodectus hesperus (black widow spider) species is renowned for its tensile strength, toughness and elasticity. These properties allow spider silk to outperform some of the best man-made materials such as Kevlar. Despite 6-7 different fiber types, our lab focuses mainly on Major Ampullate (MA) silk. In particular, we are involved in studying the structural proteins (spidroins), which are the molecules that comprise a vast amount of the silk. Previous proteome analysis of dragline silk has revealed approximately 48 proteins - most are uncharacterized. We hypothesize that our proteome analysis of dragline silk will provide a more comprehensive list of proteins, further revealing unknown proteins that are constituents of dragline silk. In order to discover new proteins, we dissolved dragline silk from black widow spiders with urea to denature the proteins. Proteins were further unfolded using reducing agent, followed by alkylation of cysteine sidechain groups. Following these steps, the proteins were digested with trypsin and the peptides separated using nano high performance liquid chromatography. Peptides were subject to MS and MS/MS analysis using an Orbitrap Fusion mass spectrometer. Spectra were analyzed using Proteome Discover 2.1 software. From our experiment we have generated a new list of proteins that more accurately depicts proteins within dragline silk.