Astacin family metallopeptidases and serine peptidase inhibitors in spider digestive fluid.
Digestive fluid of the araneid spider Argiope aurantia is known to contain zinc metallopeptidases. Using anion-exchange chromatography, size-exclusion chromatography, sucrose density gradient centrifugation, and gel electrophoresis, we isolated two lower-molecular-mass peptidases, designated p16 and p18. The N-terminal amino acid sequences of p16 (37 residues) and p18 (20 residues) are 85% identical over the first 20 residues and are most similar to the N-terminal sequences of the fully active form of meprin (beta subunits) from several vertebrates (47-52% and 50-60% identical, respectively). Meprin is a peptidase in the astacin (M12A) subfamily of the astacin (M12) family. Additionally, a 66-residue internal sequence obtained from p16 aligns with the conserved astacin subfamily domain. Thus, at least some spider digestive peptidases appear related to astacin of decapod crustaceans. However, important differences between spider and crustacean metallopeptidases with regard to isoelectric point and their susceptibility to hemolymph-borne inhibitors are demonstrated. Anomalous behavior of the lower-molecular-mass Argiope peptidases during certain fractionation procedures indicates that these peptidases may take part in reversible associations with each other or with other proteins. A. aurantia digestive fluid also contains inhibitory activity effective against insect digestive peptidases. Here we present evidence for at least thirteen, heat-stable serine peptidase inhibitors ranging in molecular mass from about 15 to 32 kDa.
Foradori
Matthew J
MJ
Department of Zoology, Rudman Hall, 46 College Rd., University of New Hampshire, Durham, 03824-2617, USA. foradori@cisunix.unh.edu
Tillinghast
Edward K
EK
Smith
J Stephen
JS
Townley
Mark A
MA
Mooney
Robert E
RE
D000595:Amino Acid Sequence; D000818:Animals; D005766:Gastrointestinal Contents / Q000201:enzymology; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D015842:Serine Proteinase Inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D013112:Spiders / Q000201:enzymology* / Q000378:metabolism
D015842:Serine Proteinase Inhibitors; D008666:Metalloendopeptidases; C071288:astacin