Amino Acid Sequences of Neuropeptides in the Sinus Gland of the Land Crab Cardisoma carnifex: A Novel Neuropeptide Proteolysis Site

The sinus gland is a major neurosecretory structure in Crustacea. Five peptides, labeled C, D, E, F, and I, isolated from the sinus gland of the land crab have been hypothesized to arise from the incomplete proteolysis at two internal sites on a single biosynthetic intermediate peptide “H,” based on amino acid composition additivities and pulse‐chase radiolabeling studies. The presence of only a single major precursor for the sinus gland peptides implies that peptide H may be synthesized on a common precursor with crustacean hyperglycemic hormone forms, “J” and “L,” and a peptide, “K,” similar to peptides with molt inhibiting activity. Here I report the amino acid sequences of these peptides. The amino terminal sequence of the parent peptide, H, (and the homologous fragments) proved refractory to Edman degradation. Data from amino acid analysis and carboxypeptidase digestion of the naturally occurring fragments and of fragments produced by endopeptidase digestion were used together with Edman degradation to obtain the sequences. Amino acid analysis of fragments of the naturally occurring “overlap” peptides (those produced by internal cleavage at one site on H) was used to obtain the sequences across the cleavage sites. The amino acid sequence of the land crab peptide H is Arg‐Ser‐Ala‐Asp‐Gly‐Phe‐Gly‐Arg‐Met‐Glu‐Ser‐Leu‐Leu‐Thr‐Ser‐Leu‐Arg‐Gly‐Ser‐Ala‐Glu‐Ser‐Pro‐Ala‐Ala‐Leu‐Gly‐Glu‐Ala‐Ser‐Ala‐Ala‐His‐Pro‐Leu‐Glu. In vivo cleavage at one site involves excision of arginine from the sequence Leu‐Arg‐Gly, whereas cleavage at the other site involves excision of serine from the sequence Glu‐Ser‐Leu. Proteolysis at the latter sequence has not been previously reported in intact secretory granules. The aspartate at position 4 is possibly covalently modified.