The alkaline proteases of Argiope—II. Fractionation of protease activity and isolation of a silk fibroin digesting protease

Kavanagh, Edward; Tillinghast, Edward K.

doi:10.1016/0305-0491(83)90026-3

Cited by 7 publications

(8 citation statements)

References 20 publications

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“…He suspected that he had isolated just a single peptidase and that reversible association accounted for its unexpected elution pattern. We believe that Mebs' hypothesis provides the best explanation for our data and also explains why previous efforts to isolate SDF peptidases have resulted in low yields (Kavanagh and Tillinghast 1983). Future efforts may benefit from finding conditions of pH, ionic environment, and temperature that favor the dissociation of the peptidases prior to and during fractionation.…”

Section: Sdf Peptidasesmentioning

confidence: 52%

“…If, as suggested above, the spider venom peptidases were actually from SDF, then subsequent observations have confirmed the relevance of this comparison. Like astacin, the principal spider digestive peptidases are zinc metallopeptidases (Kavanagh 1979;Kavanagh and Tillinghast 1983;Foradori et al 2001). Moreover, in this study, database searches of two extended sequences from p16 and the N-terminus of p18 demonstrated that these peptidases belong to the astacin (M12A) subfamily of the astacin (M12) family (Rawlings et al 2004, merops.sanger.ac.uk).…”

Section: Sdf Peptidasesmentioning

confidence: 64%

“…Like astacin, meprin A has a considerably lower pI (4-5) (Butler et al 1987) than the bulk of the peptidases in SDF. It remains to be established which peptidases, by the designations used in this paper, correspond to the four Argiope peptidase fractions (A-D) of Kavanagh (1979) and Kavanagh and Tillinghast (1983), and in particular to the apparently homogeneous Argiope protease B and Argiope protease D. Similarities with respect to molecular mass, pI, and relative abundance make it likely that p16 is one of these two peptidases. It also remains to be established if, as with many other enzymes including astacin (Zwilling and Neurath 1981;Stöcker et al 1991b), α-amylase (Mommsen 1978b,d), and Nacetyl-β-glucosaminidase (Mommsen 1978b), any of the more minor Argiope peptidases (observed in SDS-PAGE gels in the vicinity of p35, p28, and p16 (Figs.…”

Section: Sdf Peptidasesmentioning

confidence: 95%

“…The digestive peptidases of orb-weaving spiders are necessary not only for consumption of prey, but also for the spider's recycling of its old orb webs into new (Tillinghast and Kavanagh 1977;Kavanagh and Tillinghast 1983;Townley and Tillinghast 1988). For some orb-weavers, including the araneid Argiope aurantia Lucas 1833, peptidases also play a role in the emergence of young spiders from the egg sac (Foradori et al 2002).…”

Section: Introductionmentioning

confidence: 99%

“…The most comprehensive studies to date are those of Mommsen (1978aMommsen ( ,b,c,d, 1980. In earlier studies (Tillinghast and Kavanagh 1977;Kavanagh 1979;Kavanagh and Tillinghast 1983), two peptidases were isolated from the digestive fluid of A. aurantia (though additional peptidases were demonstrated and partially characterized). Designated Argiope protease B and Argiope protease D, they had estimated molecular masses of 17.6 and 20.2 kDa, respectively, slightly alkaline pH optima, were cationic at physiological pH (about 7.8), and were shown to be metallopeptidases.…”

Section: Introductionmentioning

confidence: 99%

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Astacin family metallopeptidases and serine peptidase inhibitors in spider digestive fluid

Foradori

Tillinghast

Smith

et al. 2006

Comparative Biochemistry and Physiology Part B: Biochemistry an

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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 (β 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.

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Section: Sdf Peptidasesmentioning

confidence: 52%

Section: Sdf Peptidasesmentioning

confidence: 64%