Authentic FMRFamide and two FMRFamide-related heptapeptides were purified from the central nervous system of the fresh water snail Lymnaea stagnalis. The sequences of the heptapeptides were determined as: Ser-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (SDPFLRFamide) and Gly-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (GDPFLRFamide) by modified Edman degradation and enzymatic digestion. Relatively high quantities of the deamidated and therefore non-immunoreactive analogs of these two peptides (SDPFLRF and GDPFLRF) were also found. SDPFLRFamide and GDPFLRFamide were synthesized and were found to be chromatographically and biologically indistinguishable from the natural peptides, confirming the sequences. The log dose-response curves for the chronotropic action of either synthetic peptide on the heart of Lymnaea was very similar to that of FMRFamide. These data indicate that Lymnaea contains a family of FMRFamide-like peptides.

Amino Acid Sequence↗

PubMedDate not supplied

Evidence for a novel FMRFamide-related heptapeptide in the pulmonate snail Siphonaria pectinata.

Extracts of whole false limpets (Siphonaria pectinata) were analysed to determine their complement of FMRFamide-related peptides. As in other pulmonates, FMRFamide itself was found to account for only a portion of the immunoreactivity; the largest immunoreactive peptide peak eluted during HPLC under acidic conditions at the same position as a peak also found in other pulmonates. This major peak was resolved into two components by HPLC at neutral pH, and one component was identified as the heptapeptide amide, GDPFLRFamide, previously described from Lymnaea. The amino acid composition of the second component indicates that it is also a heptapeptide, but that it has two Asx (aspartic acid or asparaginyl) residues instead of the one found in the previously identified pulmonate heptapeptides.

Amino Acids↗

PubMedDate not supplied

Further characterization of Helix FMRFamide receptors: kinetics, tissue distribution, and interactions with the endogenous heptapeptides.

The biphasic binding of 125I-daYFnLRFamide to crude brain membranes of Helix aspersa is due to two discernible sites (high and low affinity) rather than different agonist-induced states. The tissues in the snail that show the greatest specific 125I-daYFnLRFamide binding are the brain, reproductive system, and digestive system. The heart shows moderate binding levels, whereas low values are obtained in the oviduct and retractor muscles. The N-terminal SAR of the Helix heptapeptides (X-DPFLRFamide) indicates that, although the substitution of Leu for Met accounts for some, the dipeptide X-Asp produces most of the loss in potency at FMRFamide receptors in Helix brain.

Amino Acid Sequence↗

PubMedDate not supplied

Authentic FMRFamide is present in the polychaete Nereis virens.

The tetrapeptide FMRFamide is but one member of a large family of invertebrate neuropeptides which includes another tetrapeptide, FLRFamide, and several longer peptides terminating in one or the other of these tetrapeptide sequences. These peptides have been isolated from both molluscs and arthropods, but so far not one has been isolated from an annelid. Since the annelid worms are believed to share a common ancestor with molluscs and arthropods, they should contain FMRFamide-like peptides. We found two immunoreactive peaks in Nereis virens, but microsequencing and fast atom bombardment mass spectrometry revealed that they represent only one native peptide, FMRFamide. (The other peak is its methionyl sulfoxide derivative.) Each worm contained only 100 to 600 fmols of peptide, which is at least 10-100 times less than the levels in molluscs. Our identification of a tetrapeptide, and only a tetrapeptide, in this worm suggests that the tetrapeptides are the more ancient members of the family, and were probably present in the common ancestors of the annelids, arthropods, and molluscs.

Amino Acid Sequence↗

PubMedDate not supplied

Two FMRFamide-like peptides from the free-living nematode Panagrellus redivivus.

Peptides of the FXRFamide family, where X = M, I or L, are broadly distributed among invertebrates. Two such peptides were purified and sequenced from the free-living nematode, Panagrellus redivivus. Immunohistochemical techniques localized FMRFamide-like material in several regions of these organisms, including the nerve cords and, most prominently, in paired groups of cells located caudally to the base of the pharynx. RIA determinations gave an estimate of 2.8 nmol immunoreactive peptide/g of an acetone extract of P. redivivus. Four sequential HPLC purification steps, followed by sequencing by automated Edman degradation and FAB-MS, led to the identification of Ser-Asp-Pro-Asn-Phe-Leu-Arg-Phe-amide (SDPNFLRFamide) and Ser-Ala-Asp-Pro-Asn-Phe-Leu-Arg-Phe-amide (SADPNFLRFamide) as members of the FXRFamide family in this nematode.

Amino Acid Sequence↗

PubMedDate not supplied

The flp-1 propeptide is processed into multiple, highly similar FMRFamide-like peptides in Caenorhabditis elegans.

Previously, we described a gene, flp-1, that encodes seven FMRFamide-like peptides from two alternatively spliced transcripts in the nematode Caenorhabditis elegans. To determine whether all or a subset of the predicted peptides coded for by flp-1 are produced in vivo, we undertook the isolation of FMRFamide-like peptides from C. elegans. Six FLRFamide-containing peptides, all contained within the putative translation products of the flp-1 gene, were isolated from extracts of mixed stage animals. By quantitative PCR analysis of RNA from mixed stage animals, we found that the shorter transcript of flp-1 has a higher level of expression than the longer transcript.

Amino Acid Sequence↗