Characterization of a cysteine-rich protein (CRP1) found within black widow spider dragline silk
Poster Number
31
Research or Creativity Area
Natural Sciences
Abstract
Black widow dragline silk is an extraordinary fiber with high tensile strength, extensibility and toughness. Spiders use dragline silk for locomotion and web construction. Dragline silk, also known as a “safety line,” has unique mechanical properties, opening up its potential use in medicine and engineering applications. Proteomic analysis of dragline silk has identified at least eight different proteins, including major ampullate spidroin 1, 2 and 3, (MaSp1, 2, and 3), along with two cysteine-rich proteins 1 and 4 (CRP1 and CRP4). Molecular modeling predicts CRP1 contains a cysteine slipknot motif, which is hypothesized to contribute to the strength and toughness of dragline silk. In order to better understand the role of CRP1 in spider silk, we built a prokaryotic expression vector for recombinant CRP1 production in bacteria. We amplified the CRP1 cDNA using PCR, inserted it into an expression vector and expressed CRP1 in bacteria. Following expression, we purified recombinant CRP1 using affinity chromatography and confirmed its purification using mass spectrometry. Preliminary studies support that CRP1 can form larger molecular aggregates with itself, suggesting it may play a structural role in dragline silk.
Location
Don and Karen DeRosa University Center (DUC) Poster Hall
Start Date
27-4-2024 10:30 AM
End Date
27-4-2024 12:30 PM
Characterization of a cysteine-rich protein (CRP1) found within black widow spider dragline silk
Don and Karen DeRosa University Center (DUC) Poster Hall
Black widow dragline silk is an extraordinary fiber with high tensile strength, extensibility and toughness. Spiders use dragline silk for locomotion and web construction. Dragline silk, also known as a “safety line,” has unique mechanical properties, opening up its potential use in medicine and engineering applications. Proteomic analysis of dragline silk has identified at least eight different proteins, including major ampullate spidroin 1, 2 and 3, (MaSp1, 2, and 3), along with two cysteine-rich proteins 1 and 4 (CRP1 and CRP4). Molecular modeling predicts CRP1 contains a cysteine slipknot motif, which is hypothesized to contribute to the strength and toughness of dragline silk. In order to better understand the role of CRP1 in spider silk, we built a prokaryotic expression vector for recombinant CRP1 production in bacteria. We amplified the CRP1 cDNA using PCR, inserted it into an expression vector and expressed CRP1 in bacteria. Following expression, we purified recombinant CRP1 using affinity chromatography and confirmed its purification using mass spectrometry. Preliminary studies support that CRP1 can form larger molecular aggregates with itself, suggesting it may play a structural role in dragline silk.
