Spider Silk Assembly and Potential Redox Trigger

Poster Number

70

Lead Author Affiliation

Pre-dental

Lead Author Status

Undergraduate - Sophomore

Second Author Affiliation

Pre-dental

Second Author Status

Undergraduate - Sophomore

Third Author Affiliation

Pre-dental

Third Author Status

Undergraduate - Sophomore

Fourth Author Affiliation

Pre-dental

Fourth Author Status

Undergraduate - Senior

Fifth Author Affiliation

Pre-Pharmacy

Fifth Author Status

Undergraduate - Sophomore

Sixth Author Affiliation

Pre-dental

Sixth Author Status

Undergraduate - Senior

Additional Authors

7) Pre-dental, Junior

8) Pre-Pharmacy, Sophomore

9) Pre-dental, Sophomore

10) Pre-dental, Junior

11) Pre-dental, Junior

12) Pre-dental, Junior

13) Pre-dental, Junior

14) Pre-dental, Junior

15) Biological Sciences Professor

Faculty Mentor Name

Dr. Craig Vierra

Research or Creativity Area

Natural Sciences

Abstract

Spider silk, possessing incredible properties in strength, toughness, and elasticity, plays an important role in spider locomotion and web construction. Spider silk is stronger than steel and tougher than Kevlar while retaining elasticity. It has the potential to revolutionize a variety of fields, including medicine and engineering, through its synthetic replication and reproduction. Proteomic analysis of dragline silk has identified that it is predominantly composed of major ampullate spidroin proteins 1 and 2 (MaSp1 and MaSp2), along with two cysteine-rich proteins 1 and 4 (CRP1 and CRP4). MaSp1 and CRP4 are important proteins because of their contribution to the characteristic tensile strength and elastic properties of dragline silk. The trigger mechanism in the spinning duct plays an important role in the conversion of the proteins from a liquid to solid form as the silk is extruded from the spider. It is during this transformation that the unique properties of spider silk emerge. Our experiment focuses on a redox trigger as a fundamental mechanism which governs the assembly of spider silk. To test this theory, the formation of disulfide bonds between the N-terminus of MaSp1 and CRP4 were studied under different oxidation and reduction (redox) environments. Utilizing PCR, we amplified cDNAs coding for the N-terminus of MaSp1 and CRP4, and expressed these recombinant proteins in bacteria. To investigate redox as a potential trigger for spider silk assembly, proteins were purified using affinity chromatography, then analyzed by SDS-PAGE, mass spectrometry, and western blot analysis. Taken together, our findings will help advance our understanding of the biochemical mechanisms of spider silk assembly.

Location

University of the Pacific, DeRosa University Center

Start Date

26-4-2025 10:00 AM

End Date

26-4-2025 1:00 PM

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Apr 26th, 10:00 AM Apr 26th, 1:00 PM

Spider Silk Assembly and Potential Redox Trigger

University of the Pacific, DeRosa University Center

Spider silk, possessing incredible properties in strength, toughness, and elasticity, plays an important role in spider locomotion and web construction. Spider silk is stronger than steel and tougher than Kevlar while retaining elasticity. It has the potential to revolutionize a variety of fields, including medicine and engineering, through its synthetic replication and reproduction. Proteomic analysis of dragline silk has identified that it is predominantly composed of major ampullate spidroin proteins 1 and 2 (MaSp1 and MaSp2), along with two cysteine-rich proteins 1 and 4 (CRP1 and CRP4). MaSp1 and CRP4 are important proteins because of their contribution to the characteristic tensile strength and elastic properties of dragline silk. The trigger mechanism in the spinning duct plays an important role in the conversion of the proteins from a liquid to solid form as the silk is extruded from the spider. It is during this transformation that the unique properties of spider silk emerge. Our experiment focuses on a redox trigger as a fundamental mechanism which governs the assembly of spider silk. To test this theory, the formation of disulfide bonds between the N-terminus of MaSp1 and CRP4 were studied under different oxidation and reduction (redox) environments. Utilizing PCR, we amplified cDNAs coding for the N-terminus of MaSp1 and CRP4, and expressed these recombinant proteins in bacteria. To investigate redox as a potential trigger for spider silk assembly, proteins were purified using affinity chromatography, then analyzed by SDS-PAGE, mass spectrometry, and western blot analysis. Taken together, our findings will help advance our understanding of the biochemical mechanisms of spider silk assembly.