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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.
J Biol Chem. 2009 Oct 16; 284(42):29097-108.JB

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 in-solution 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.

Authors+Show Affiliations

Department of Biological Sciences, University of the Pacific, Stockton, California 95211, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

19666476

Citation

Blasingame, Eric, et al. "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." The Journal of Biological Chemistry, vol. 284, no. 42, 2009, pp. 29097-108.
Blasingame E, Tuton-Blasingame T, Larkin L, et al. 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. J Biol Chem. 2009;284(42):29097-108.
Blasingame, E., Tuton-Blasingame, T., Larkin, L., Falick, A. M., 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. The Journal of Biological Chemistry, 284(42), 29097-108. https://doi.org/10.1074/jbc.M109.021378
Blasingame E, et al. 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. J Biol Chem. 2009 Oct 16;284(42):29097-108. PubMed PMID: 19666476.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Blasingame,Eric, AU - Tuton-Blasingame,Tiffany, AU - Larkin,Leah, AU - Falick,Arnold M, AU - Zhao,Liang, AU - Fong,Justine, AU - Vaidyanathan,Veena, AU - Visperas,Anabelle, AU - Geurts,Paul, AU - Hu,Xiaoyi, AU - La Mattina,Coby, AU - Vierra,Craig, Y1 - 2009/08/07/ PY - 2009/8/12/entrez PY - 2009/8/12/pubmed PY - 2009/12/16/medline SP - 29097 EP - 108 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 284 IS - 42 N2 - 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 in-solution 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. SN - 1083-351X UR - https://www.unboundmedicine.com/medline/citation/19666476/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_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=19666476 DB - PRIME DP - Unbound Medicine ER -