Degradation of Dip-allatostatins by hemolymph from the cockroach, Diploptera punctata

Incubation of Dip-AST 7 (APSGAQRLYGFGLa) or Dip-AST 9 (GDGRLYAFGLa) (5 microM) with hemolymph for 30 min results in cleavage by a putative endopeptidase, yielding the C-terminal hexapeptide. This metabolic product is subsequently cleaved by an amastatin-sensitive aminopeptidase to yield the the C-terminal pentapeptide, as treatment with the competitive aminoexopeptidase inhibitor, amastatin, results in a significant accumulation of the C-terminal hexapeptide. Interestingly, Dip-AST 5 (DRLYSFGLa) (6 microM), which in common with Dip-AST 7 and 9 possesses Arg-Leu-Tyr, is not rapidly cleaved. However, [3H-Tyr]Dip-AST 5 at physiological concentrations (4 nM), appears to be cleaved by the same enzymes that cleave Dip-AST 7 and 9, albeit at a reduced rate. Incubation of other members of the Dip-allatostatin family with hemolymph also results in cleavage of the peptides, suggesting that there are a variety of endo- and/or exopeptidases present in the hemolymph of D. punctata.