Biomedical subjects

J M Conlon

Publications and source records attributed to J M Conlon.

At least 73 records · Page 4Linked to original sources

PubMed1999

Identification of an urotensin I-like peptide in the pituitary of the lungfish Protopterus annectens: immunocytochemical localization and biochemical characterization.

In the present study we have investigated the localization and biochemical characteristics of urotensin I (UI)-like and urotensin II (UII)-like immunoreactive peptides in the central nervous system (CNS) and pituitary of the lungfish, Protopterus annectens, by using antisera raised against UI from the white sucker Catostomus commersoni and against UII from the goby Gillichythys mirabilis. UI-like immunoreactive material was found within the melanotrope cells of the intermediate lobe of the pituitary. By contrast, no UI-immunoreactive structures were found in the brain. No UII-like peptides structurally similar to goby UII were found in the brain and pituitary of P. annectens. The UI-immunoreactive material localized in the pituitary was characterized by combining reversed-phase high-performance liquid chromatography (HPLC) analysis and radioimmunological detection. The UI-like immunoreactivity contained in a pituitary extract eluted as a single peak with a retention time intermediate between those of sucker UI and rat corticotropin-releasing factor (CRF). Control tests on adjacent sections of pituitary showed that the UI antiserum cross-reacted with the frog skin peptide sauvagine, but lungfish UI did not co-elute with synthetic sauvagine on HPLC. On the contrary, no cross-reaction was observed between the UI antiserum and CRF or alpha-melanocyte-stimulating hormone (alpha-MSH). The occurrence of an UI-like peptide in the intermediate lobe of the pituitary of P. annectens suggests that, in lungfish, this peptide may act as a classic pituitary hormone or may be involved in the control of melanotrope cell secretion.

Amino Acid Sequence↗

PubMed1999

Purification and characterization of insulin and peptides derived from proglucagon and prosomatostatin from the fruit-eating fish, the pacu Piaractus mesopotamicus.

The fruit-eating teleost fish, the pacu Piaractus mesopotamicus (Characiformes, Characidae) is classified along with the carp and the catfish in the superorder Ostariophysi. The pacu is able to survive and grow in captive conditions feeding exclusively on carbohydrates. Hormonal polypeptides in an extract of pacu Brockmann bodies were purified to homogeneity by reversed phase HPLC and their primary structures determined by automated Edman degradation. Pacu insulin contains only two substitutions, Glu-->Asp at A15 and Thr-->Ser at B24 (corresponding to B22 in mammalian insulins) compared with carp insulin. The B-chains of both insulins contain a dipeptide extension to the N-terminus and a deletion of the C-terminal residue compared with human insulin. Pacu glucagon differs from catfish glucagon by a single substitution at position 17 (Arg-->Gln. The primary structure of the 34 amino acid residue glucagon-like peptide (GLP) differs from catfish GLP only at positions 12 (Ser-->Ala) and 33 (Pro-->Gln). In common with other teleost species, the pacu expresses two somatostatin genes. Somatostatin-14, derived from preprosomatostatin-I (PSS-I), is identical to mammalian/catfish somatostatin-14. Although pacu somatostatin-II was not identified in this study, a peptide was purified that shows 67% sequence identity with residues (1-58) of catfish preprosomatostatin-II (PSS-II). This relatively high degree of sequence similarity contrasts with the fact that catfish PSS-II shows virtually no sequence identity with the corresponding PSS-II from anglerfish (Acanthopterygii) and trout (Protoacanthopterygii). A comparison of the primary structures of the islet hormones suggest that amino acid sequences may have been better conserved within the Ostariophysi than in other groups of the taxon Euteleostei that have been studied.

Amino Acid Sequence↗

PubMed1999

Peptides with antimicrobial activity of the brevinin-1 family isolated from skin secretions of the southern leopard frog, Rana sphenocephala.

Three peptides with growth-inhibitory activity towards the gram-negative bacterium Eschericia coli were isolated from electrically stimulated secretions from the skin of the southern leopard frog, Rana sphenocephala. Structural characterization demonstrated that the peptides [brevinin-1Sa, minimum inhibitory concentration (MIC) = 55 microM; brevinin-1Sb, MIC = 17 microM; brevinin-1Sc, MIC = 14 microM] represent new members of the brevinin-1 family of antimicrobial peptides, previously isolated from several other species of frogs of the genus Rana. Their high concentration in skin secretions and extreme variability in amino acid sequence suggest that the brevinin family of peptides may be of value as molecular markers for the identification and taxonomic classification of Ranid frogs.

Amino Acid Sequence↗

PubMed1999

Purification, structural characterization, and myotropic activity of endothelin from trout, Oncorhynchus mykiss.

Endothelin (ET) from a nontetrapod species has never been characterized, either structurally or biologically. A single molecular form of trout ET with 21-amino-acid residues was isolated in pure form from an extract of the kidney of the steelhead trout, Oncorhynchus mykiss and its primary structure established as Cys-Ser-Cys-Ala-Thr-Phe-Leu-Asp-Lys-Glu10-Cys-Val-Tyr-Phe-Cys-His- L eu-Asp-Ile-Ile20-Trp. This amino acid sequence shows only three substitutions (Ala4-->Ser, Thr5-->Ser, and Phe6-->Trp) compared with human ET-2, demonstrating that the structure of the peptide has been well conserved during evolution and that the pathway of posttranslational processing of preproendothelin in the trout is probably similar to that in mammals. Synthetic trout ET produced concentration-dependent constrictions of isolated rings of vascular tissue from trout efferent branchial artery (EBA; pD2 = 7. 90 +/- 0.06, n = 5), caeliacomesenteric artery (pD2 = 8.03 +/- 0. 04, n = 4), anterior cardinal vein (ACV; pD2 = 8.57 +/- 0.25, n = 4), and rat abdominal aorta (AO; pD2 = 8.86 +/- 0.08, n = 7). Trout and rat vessels were more sensitive to mammalian ET-1 than to trout ET (pD(2) for human ET-1 in: EBA = 9.12 +/- 0.14; ACV = 9.90 +/- 0.15; AO = 8.86 +/- 0.08), but there was no significant difference in the maximum tension produced by either peptide in these vessels.

Amino Acid Sequence↗

PubMed1998

Antimicrobial peptides of the brevinin-2 family isolated from gastric tissue of the frog, Rana esculenta.

Four structurally related peptides with potent growth-inhibitory activity towards Escherichia coli were isolated from an extract of the stomach of the European green frog Rana esculenta, and were identified as members of the brevinin-2 family. Two peptides, termed brevinin-2Eg (GIMDTLKNLA10 KTAGKGALQS20 LLNHASCK LS30GQC) and brevinin-2Eh (GIMDTLKNLA10 KTAGKGALQS20 LLNHASCKL S30 KQC) have not been described previously. One peptide is identical to brevinin-2Ec, previously isolated from R. esculenta skin secretions, and one peptide is identical to brevinin-2Ef whose structure has been deduced from a cloned cDNA prepared from a R. esculenta skin cDNA library. The data demonstrate that certain peptides of the brevinin-2 family, like the magainins in the toad, Xenopus laevis, may play an important role in protecting the gastrointestinal tract of Ranid frogs against microbial invasion.

Amino Acid Sequence↗

PubMed1998

Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord.

Urotensin II (UII) is a cyclic peptide initially isolated from the caudal neurosecretory system of teleost fish. Subsequently, UII has been characterized from a frog brain extract, indicating that a gene encoding a UII precursor is also present in the genome of a tetrapod. Here, we report the characterization of the cDNAs encoding frog and human UII precursors and the localization of the corresponding mRNAs. In both frog and human, the UII sequence is located at the C-terminal position of the precursor. Human UII is composed of only 11 amino acid residues, while fish and frog UII possess 12 and 13 amino acid residues, respectively. The cyclic region of UII, which is responsible for the biological activity of the peptide, has been fully conserved from fish to human. Northern blot and dot blot analysis revealed that UII precursor mRNAs are found predominantly in the frog and human spinal cord. In situ hybridization studies showed that the UII precursor gene is actively expressed in motoneurons. The present study demonstrates that UII, which has long been regarded as a peptide exclusively produced by the urophysis of teleost fish, is actually present in the brain of amphibians and mammals. The fact that evolutionary pressure has acted to conserve fully the biologically active sequence of UII suggests that the peptide may exert important physiological functions in humans.

Amino Acid Sequence↗

PubMed1998

Comparative vascular responses in elasmobranchs to different structures of neuropeptide Y and peptide YY.

The vascular responses to neuropeptide Y (NPY) and peptide YY (PYY) were tested in several species of elasmobranchs to assess whether changes in sequence in these neuropeptides from elasmobranchs to mammals are associated with different physiological responses. NPY-like immunoreactivity was detected in the gut and in nerve fibres surrounding some, but not all, blood vessels of six species. Intravenous injection of dogfish, frog and human NPY in anaesthetised fish caused similar vasopressor effects in the three species tested, except human NPY which lowered blood pressure in one of the three. Dogfish NPY and PYY were equipotent pressor agents in two species, but PYY was significantly more potent than NPY in one species. NPY and PYY both contracted isolated gut arteries from three species, but had no effect on isolated efferent arteries tested. In conclusion, differential vascular responses in elasmobranchs are not associated with changes in NPY sequence across vertebrates, but may be with changes in PYY in some species.

Amino Acid Sequence↗

PubMed1998

Amino terminal glycation of gastric inhibitory polypeptide enhances its insulinotropic action on clonal pancreatic B-cells.

Gastric inhibitory polypeptide (GIP) is a potent insulin-releasing hormone of the enteroinsular axis. This study has examined glycation of GIP and effects of such structural modification on insulin secretion from a glucose-responsive clonal pancreatic B-cell line (BRIN-BD11). Monoglycated GIP (Mr 5149.5) was prepared by incubation with d-glucose under reducing conditions and purified by HPLC. Automated Edman degradation and mass spectrometric analysis indicated that GIP was specifically glycated at the amino terminus. In acute (20 min) incubations at 5.6 mM glucose, GIP (3x10-11-10-8 M) significantly stimulated insulin secretion by 1.6-2.1-fold from BRIN-BD11 cells. The stimulatory effect induced by GIP over this concentration range was further enhanced by 1.5-2.5-fold following N-terminal glycation. These data indicate that GIP can be glycated under hyperglycaemic conditions at the amino terminal Tyr1, and that this modification increases the glucose-dependent insulinotropic action of the peptide.

Amino Acids↗

PubMed1998

Ranatuerins: antimicrobial peptides isolated from the skin of the American bullfrog, Rana catesbeiana.

Nine peptides, termed ranatuerins 1-9, with antimicrobial activity towards Staphylococcus aureus, were isolated from an extract of the skin of the adult American bullfrog, Rana catesbeiana. In common with other cytolytic peptides from Ranid frogs, (e.g. ranalexin, gaegurins, brevinins), ranatuerins 1 and 4 contain an intramolecular disulfide bridge forming a heptapeptide ring whereas in ranatuerins 2 and 3 the disulfide bridge forms a hexapeptide ring. The structurally related ranatuerins 5-9 comprise 12 - 14 amino acids and show sequence similarity towards the hemolytic peptides A1 and B9 previously isolated from the skin of Rana esculenta. Of the peptides purified, ranatuerin 1 (SMLSVLKNLGKVGLG FVACKINKQC) showed the broadest spectrum of antimicrobial action with inhibitory activity against S. aureus, Escherichia coli and Candida albicans.

Amino Acid Sequence↗

PubMed1998