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.
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.
Blasingame
Eric
E
Department of Biological Sciences, University of the Pacific, Stockton, California 95211, USA.
Tuton-Blasingame
Tiffany
T
Larkin
Leah
L
Falick
Arnold M
AM
Zhao
Liang
L
Fong
Justine
J
Vaidyanathan
Veena
V
Visperas
Anabelle
A
Geurts
Paul
P
Hu
Xiaoyi
X
La Mattina
Coby
C
Vierra
Craig
C
D000818:Animals; D001740:Black Widow Spider; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D005349:Fibroins / Q000737:chemistry*; D015723:Gene Library; D013058:Mass Spectrometry / Q000379:methods*; D008855:Microscopy, Electron, Scanning / Q000379:methods; D010455:Peptides / Q000737:chemistry; D017434:Protein Structure, Tertiary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D047011:Silk / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013112:Spiders; D014357:Trypsin / Q000737:chemistry
D018076:DNA, Complementary; D010455:Peptides; D047011:Silk; C086087:spidroin 1; D005349:Fibroins; D014357:Trypsin