The peptide with structural similarity to mammalian substance P (M-SP) has been isolated from extract of the body of the African lungfish, Protopterus dolloi, using the rectum of the newt as the bioassay system. The primary structure of the SP-related peptide was identified as Lys-Pro-Arg-Pro-Asp-Gln-Phe-Tyr-Gly-Leu-Met-NH2 (L-SP) and contained four substitutions ($Lys^{1} ightarrow $ Arg, $Arg^{3} ightarrow$ Lys, $Asp^{5} ightarrow$ Gln, and $Tyr^{8} ightarrow$ Phe) compared with M-SP; this structure is identical to that of the peptide isolated from the gut of the Australian lungfish. Circular dichroism spectra showed that L-SP had an unordered structure in the buffer solution and phospholipid bilayers. This peptide was found to have an excitatory effect on rectal muscle tissues of newt, quail, and fish. L-SP also had a more potent vasodilatory effect on the guinea-pig aorta than that of M-SP. The identification of the peptide provides evidence that SP family, hitherto confined to mammals, have a widespread occurrence in lungfish.