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Molecular identification of silk proteins in the gumfooted lines and attachment discs of the black widow spider, latrodectus hesperus

Author

Eric M. Blasingame, University of the Pacific

Date of Award

2009

Document Type

Thesis - Pacific Access Restricted

Degree Name

Master of Science (M.S.)

Department

Biological Sciences

First Advisor

Craig A. Vierra

First Committee Member

Patrick R. Jones

Second Committee Member

Andreas Franz

Third Committee Member

Geoff Lin-Cereghino

Abstract

Silks from araneoid spiders have become an active area of research for material scientists, biochemists, and molecular biologists. Mechanical properties of spider silk such as elasticity, tensile strength, and toughness make the manufacturing of silk for medical sutures, body armor, ropes and other synthetic material applications great possibilities. The difficulties of having a black widow spider farm to harvest silk, due to their cannibalistic nature, make recombinant expression of silk proteins a fundamental goal of spider silk research. In order to express silk fibers, cDNAs encoding the corresponding silk fiber products must first be isolated and identified. One of the first steps in gene identification relies on the identification of the proteins in the silk fibers.

No previous study has demonstrated the molecular constituents of gumfooted lines. In the course of this research, the core fibroins in the gumfooted lines were identified to be members of the Major Ampullate Spidroin family (MaSp), using mass spectrometry. This research was the first to identify the core fibroins of the gumfooted lines.

Novel peptide fragments from solubilized gumfooted lines were acquired from manual de novo MSIMS sequencing after in-gel tryptic digestion. These peptide fragments showed post-translational modifications consistent with glycosylation, which aligns with the reported chemical properties of glue proteins.

Novel peptide sequences were also acquired from the attachment discs as well as novel scanning electron microscopy images and reveal, for the first time, the physical attributes and molecular properties of threads attached to the surface of an immobilized structure. This study was the first to identify the molecular constituents of the attachment discs.

Pages

79

Recommended Citation

Blasingame, Eric M.. (2009). Molecular identification of silk proteins in the gumfooted lines and attachment discs of the black widow spider, latrodectus hesperus. University of the Pacific, Thesis - Pacific Access Restricted. https://scholarlycommons.pacific.edu/uop_etds/731