Finding of primitive polyamine toxins in the venom of a joro spider, Nephila clavata.

Abstract

A series of joro spider toxins, novel polyamines sharing a common moiety of 2,4-dihydroxyphenylacetyl cadaverine, have been identified using various bioassays, such as inhibition of a glutamatergic transmission and insecticidal activity. In this paper, we tried to chemically find still unknown polyamine toxins in the venom of a joro spider, N. clavata, by several analytical methods based on the characteristics of the common moiety. An aqueous extract from 3000 venoms was separated by preparative high performance liquid chromatography (HPLC). The polyamine toxins were detected by monitoring the fluorescence produced in an on-line reaction of o-phthalaldehyde with amino groups and UV absorption of the phenol group. Two compounds in minute quantity were purified and analyzed by gas-liquid chromatography (GC) and HPLC, which we specifically developed for the simultaneous determination of amino acids and polyamines of the toxins. Judging from the constituents of the hydrolysate by GC and HPLC and the molecular weights determined by fast atom bombardment mass spectrometry, the two compounds were estimated to be N-(2,4-dihydroxyphenylacetyl-L-asparaginyl)-N'- (3-aminopropyl-beta-alanyl) cadaverine and N-(4-hydroxyphenylacetyl-L-asparaginyl)-N'-(3-aminopropyl-beta-ala nyl) cadaverine. These compounds were small in content and molecular weight compared with hitherto known toxins. Both were presumed to be biochemically primitive toxins and were named spidamine and joramine, respectively.