Peptides from the venom ducts of cone snails (genus Coitus) contain gamma-carboxyglutamate residues. The gamma-glutamyl carboxylase responsible for this post-translational modification is localized in the microsomal fraction, strictly dependent on vitamin It, activated by ammonium sulfate, and is associated with endogenous substrate. The K-m of the enzyme for vitamin K is comparable to that for the bovine carboxylase, However, a propeptide containing substrate related to the blood coagulation protein factor IX, a highly efficient substrate for the bovine enzyme, was poorly carboxylated by the Conus enzyme, suggesting differences in gamma-carboxylase recognition signal sequences and/or structural requirements at the carboxylation site. (C) 1997 Federation of European Biochemical Societies.