Pyriform spidroin 1, a novel member of the silk gene family that anchors dragline silk fibers in attachment discs of the black widow spider, Latrodectus hesperus

Eric Blasingame, Tiffany Tuton-Blasingame, Leah Larkin, Arnold M. Falick, Liang Zhao, Justine Fong, Veena Vaidyanathan, Anabelle Visperas, Paul Geurts, Xiaoyi Hu, Coby La Mattina, Craig Vierra

Research output: Contribution to journalArticle

Abstract

Spiders spin high performance threads that have diverse mechanical properties for specific biological applications. To better understand the molecular mechanism by which spiders anchor their threads to a solid support, we solubilized the attachment discs from black widow spiders and performed insolution tryptic digests followed by MS/MS analysis to identify novel peptides derived from glue silks. Combining matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry and cDNA library screening, we isolated a novel member of the silk gene family called pysp1 and demonstrate that its protein product is assembled into the attachment disc silks. Alignment of the PySp1 amino acid sequence to other fibroins revealed conservation in the non-repetitive C-terminal region of the silk family. MS/MS analysis also confirmed the presence of MaSp1 and MaSp2, two important components of dragline silks, anchored within the attachment disc materials. Characterization of the ultrastructure of attachment discs using scanning electron microscopy studies support the localization of PySp1 to small diameter fibers embedded in a glue-like cement, which network with large diameter dragline silk threads, producing a strong, adhesive material. Consistent with elevated PySp1 mRNA levels detected in the pyriform gland, MS analysis of the luminal contents extracted from the pyriform gland after tryptic digestion support the assertion that PySp1 represents one of the major constituents manufactured in the pyriform gland. Taken together, our data demonstrate that PySp1 is spun into attachment disc silks to help affix dragline fibers to substrates, a critical function during spider web construction for prey capture and locomotion.

Original languageEnglish (US)
Pages (from-to)29097-29108
Number of pages12
JournalJournal of Biological Chemistry
Volume284
Issue number42
DOIs
StatePublished - Oct 16 2009
Externally publishedYes

Fingerprint

Black Widow Spider
Silk
Anchors
Genes
Fibers
Spiders
Adhesives
Glues
Fibroins
Locomotion
spidroin 1
Gene Library
Electron Scanning Microscopy
Ionization
Mass spectrometry
Digestion
Amino Acid Sequence
Desorption
Conservation
Mass Spectrometry

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Pyriform spidroin 1, a novel member of the silk gene family that anchors dragline silk fibers in attachment discs of the black widow spider, Latrodectus hesperus. / Blasingame, Eric; Tuton-Blasingame, Tiffany; Larkin, Leah; Falick, Arnold M.; Zhao, Liang; Fong, Justine; Vaidyanathan, Veena; Visperas, Anabelle; Geurts, Paul; Hu, Xiaoyi; La Mattina, Coby; Vierra, Craig.

In: Journal of Biological Chemistry, Vol. 284, No. 42, 16.10.2009, p. 29097-29108.

Research output: Contribution to journalArticle

Blasingame, E, Tuton-Blasingame, T, Larkin, L, Falick, AM, Zhao, L, Fong, J, Vaidyanathan, V, Visperas, A, Geurts, P, Hu, X, La Mattina, C & Vierra, C 2009, 'Pyriform spidroin 1, a novel member of the silk gene family that anchors dragline silk fibers in attachment discs of the black widow spider, Latrodectus hesperus', Journal of Biological Chemistry, vol. 284, no. 42, pp. 29097-29108. https://doi.org/10.1074/jbc.M109.021378
Blasingame, Eric ; Tuton-Blasingame, Tiffany ; Larkin, Leah ; Falick, Arnold M. ; Zhao, Liang ; Fong, Justine ; Vaidyanathan, Veena ; Visperas, Anabelle ; Geurts, Paul ; Hu, Xiaoyi ; La Mattina, Coby ; Vierra, Craig. / Pyriform spidroin 1, a novel member of the silk gene family that anchors dragline silk fibers in attachment discs of the black widow spider, Latrodectus hesperus. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 42. pp. 29097-29108.
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abstract = "Spiders spin high performance threads that have diverse mechanical properties for specific biological applications. To better understand the molecular mechanism by which spiders anchor their threads to a solid support, we solubilized the attachment discs from black widow spiders and performed insolution tryptic digests followed by MS/MS analysis to identify novel peptides derived from glue silks. Combining matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry and cDNA library screening, we isolated a novel member of the silk gene family called pysp1 and demonstrate that its protein product is assembled into the attachment disc silks. Alignment of the PySp1 amino acid sequence to other fibroins revealed conservation in the non-repetitive C-terminal region of the silk family. MS/MS analysis also confirmed the presence of MaSp1 and MaSp2, two important components of dragline silks, anchored within the attachment disc materials. Characterization of the ultrastructure of attachment discs using scanning electron microscopy studies support the localization of PySp1 to small diameter fibers embedded in a glue-like cement, which network with large diameter dragline silk threads, producing a strong, adhesive material. Consistent with elevated PySp1 mRNA levels detected in the pyriform gland, MS analysis of the luminal contents extracted from the pyriform gland after tryptic digestion support the assertion that PySp1 represents one of the major constituents manufactured in the pyriform gland. Taken together, our data demonstrate that PySp1 is spun into attachment disc silks to help affix dragline fibers to substrates, a critical function during spider web construction for prey capture and locomotion.",
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