Analysis of aqueous glue coating proteins on the silk fibers of the cob weaver, Latrodectus hesperus.
Elucidation of the molecular composition and physical properties of spider glue is necessary to understand its function in the mechanics of the web and prey capture. Previous reports have indicated that components of the adhesive coating contain inorganic molecules, phosphorylated glycoproteins, lipids, and organic low-molecular mass (LMM) compounds. Using a proteomic strategy, we have investigated the viscid, aqueous components that coat different silk fiber types from the black widow spider, Latrodectus hesperus. After in-solution tryptic digestion of the aqueous protein material extracted from egg case sacs, gumfooted lines, and the web scaffolding connection joints, followed by peptide analysis using MALDI tandem TOF mass spectrometry, we demonstrate that these fibers are coated with common peptides. Utilizing a reverse genetics approach, we have isolated the cDNAs encoding two distinct fiber coating products, which we have named spider coating peptide 1 and 2 (SCP-1 and SCP-2). Secreted forms of SCP-1 and SCP-2 contain 36 and 19 amino acids, respectively, and their primary sequences display no significant similarities to ensemble repeat units from traditional fibroins. Quantitative real-time reverse transcription PCR analyses show that these mRNAs are chiefly produced by the aggregate gland. Biochemical studies also demonstrate that the SCP-1 peptide has intrinsic metal binding properties, suggesting a role of peptide-metal ion interactions with the fiber constituents to enhance thread performance. Collectively, these investigations are the first to reveal a novel role for the aggregate gland in the production of peptides that coat spider silk threads.
Hu
Xiaoyi
X
Department of Chemistry, University of the Pacific, Stockton, California 95211, USA.
Yuan
Jing
J
Wang
Xiaodong
X
Vasanthavada
Keshav
K
Falick
Arnold M
AM
Jones
Patrick R
PR
La Mattina
Coby
C
Vierra
Craig A
CA
D000269:Adhesives / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D001740:Black Widow Spider / Q000235:genetics; D005349:Fibroins; D019476:Insect Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D010063:Ovum / Q000737:chemistry; D016415:Sequence Alignment; D047011:Silk / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000269:Adhesives; D019476:Insect Proteins; D047011:Silk; D005349:Fibroins