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1.
Somatostatin-14 (SRIF-14) has been shown to occur throughout the male rat reproductive system by SRIF radioimmunoassay, Sephadex G-50 exclusion chromatography, and parallel line analysis. SRIF-28 was found only in the epididymis. The highest concentrations of total SRIF-like immunoreactivity (SLI), representing the combined concentrations of SRIF-14 and SRIF-28, were measured in the prostates of 1 1/2- and 3-month-old Sprague-Dawley rats. The levels of SLI in prostates from 9-month-old males were about 10% that of the younger animals. Dilution curves for extracts of all reproductive tissues were parallel with synthetic SRIF-14. 相似文献
2.
D LeRoith W Pickens L K Crosby M Berelowitz M Holtgrefe J Shiloach 《Biochimica et biophysica acta》1985,838(3):335-342
Extracts of Escherichia coli grown in defined medium contain somatostatin-related material (1-10 pg/g wet weight of cells). Preconditioned medium had no immunoactive somatostatin whereas, conditioned medium had 110-150 pg/l. Following purification of the extracted material on Sep-pak C18, Bio-Gel P-6 and HPLC, multiple molecular weight forms of somatostatin- (SRIF-) related material were identified. The material in one peak reacted in both the N-terminal and C-terminal SRIF immunoassay and coeluted on HPLC with SRIF-28, whereas that in a second peak eluted near SRIF-14 and was reactive only in the C-terminal SRIF assay. The two peaks are thus similar to SRIF-28 and SRIF-14 of vertebrates. These findings add support to the suggestion that vertebrate-type peptide hormones and neuropeptides have early evolutionary origins. 相似文献
3.
D. LeRoith W. Pickens L.K. Crosby M. Berelowitz M. Holtgrefe J. Shiloach 《Biochimica et Biophysica Acta (BBA)/General Subjects》1985,838(3):335-342
Extracts of Escherichia coli grown in defined medium contain somatostatin-related material (1–10 pg/g wet weight of cells). Preconditioned medium had no immunoactive somatostatin whereas, conditioned medium had 110–150 pg/1. Following purification of the extracted material on Sep-pak C18, Bio-Gel P-6 and HPLC, multiple molecular weight forms of somatostatin- (SRIF-) related material were identified. The material in one peak reacted in both the N-terminal and C-terminal SRIF immunoassay and coeluted on HPLC with SRIF-28, whereas that in a second peak eluted near SRIF-14 and was reactive only in the C-terminal SRIF assay. The two peaks are thus similar to SRIF-28 and SRIF-14 of vertebrates. These findings add support to the suggestion that vertebrate-type peptide hormones and neuropeptides have early evolutionary origins. 相似文献
4.
The effect of various vertebrate somatostatins (SRIF) on basal growth hormone (GH) secretion from goldfish pituitary fragments was studied using an in vitro perifusion system. SRIF-14 caused a rapid and dose-dependent decrease in the rate of GH release from goldfish pituitary fragments. The half-maximal effective dose (ED50) of SRIF-14 was calculated as 1.3 nM following exposure to two minute pulses of increasing concentrations of SRIF-14, whereas the ED50 of SRIF-14 calculated after continuous exposure to sequentially increasing doses of SRIF-14 was 65 nM. This difference suggests that the pituitary fragments were less responsive to SRIF-14 in the latter experiment, possibly as a result of previous exposure to SRIF-14. SRIF-28 was found to be equipotent with SRIF-14 in decreasing basal GH secretion from the goldfish pituitary. In contrast, catfish SRIF-22, a uniquely teleost SRIF isolated from catfish pancreatic islets, did not alter GH secretion. These results provide further support for the hypothesis that SRIF-14 or a very similar molecule functions as a GH release-inhibiting factor in teleosts, indicating that this action of SRIF-14 has been fully conserved throughout vertebrate evolution. 相似文献
5.
Y Bourbonnais A Danoff D Y Thomas D Shields 《The Journal of biological chemistry》1991,266(20):13203-13209
The peptide somatostatin (SRIF) exists as two different molecular species. In addition to the most common form, which is a 14-residue peptide, there is also a 14-amino acid amino-terminally extended form of the tetradecapeptide, SRIF-28. Both peptides are synthesized as larger precursors containing paired basic and monobasic amino acids at their processing sites, which, upon cleavage, generate either SRIF-14 or -28, respectively. In mammals a single prepro-SRIF molecule undergoes tissue-specific processing to generate the mature hormone whereas in some species of fish separate genes encode two distinct but homologous precursors prepro-SRIF-I and -II that give rise to SRIF-14 and -28, respectively. To investigate the molecular basis for differential processing of the prohormones we introduce their cDNAs into yeast cells (Saccharomyces cerevisiae). The signal peptides of both precursors were poorly recognized by the yeast endoplasmic reticulum translocation apparatus, consequently only low levels of SRIF peptides were synthesized. To circumvent this problem a chimeric precursor consisting of the alpha-factor signal peptide plus 30 residues of the proregion was fused to pro-SRIF-II. This fusion protein was efficiently transported through the yeast secretory pathway and processed to SRIF-28 exclusively, which is identical to the processing of the native precursor in pancreatic islet D-cells. Most significantly, cleavage of the precursor to SRIF-28 was independent of the Kex 2 endoprotease since processing occurred efficiently in a kex 2 mutant strain. We conclude that in addition to the Kex 2 protease, yeast possess a distinct prohormone converting enzyme with specificity toward monobasic processing sites. 相似文献
6.
E Moyse R Benoit A Enjalbert J P Gautron C Kordon N Ling J Epelbaum 《Regulatory peptides》1984,9(1-2):129-137
Subcellular distribution and binding characteristics of the three endogenous peptides somatostatin-14 (SRIF-14), somatostatin-28 (SRIF-28) and somatostatin-28(1-12) (SRIF-28(1-12] derived from preprosomatostatin were investigated in the rat brain cortex. The three peptides are predominantly recovered from a crude mitochondrial pellet (P2), containing the pinched off nerve endings. Specific high affinity binding sites for 125I-N-Tyr-SRIF-14 and 125I-N-Tyr-SRIF-28 are present on pituitary and brain membranes. Under the same conditions, 125I-N-Tyr-SRIF-28(1-12) binding is undetectable. Moreover, SRIF-28(1-12) does not displace 125I-N-Tyr-SRIF-14 or 125I-N-Tyr-SRIF-28 binding. SRIF-28 is more potent than SRIF-14 to displace 125I-N-Tyr-SRIF-28 binding to brain and pituitary membranes, while both peptides are equipotent to displace 125I-N-Tyr-SRIF-14 binding. Finally, the regional distribution of 125I-N-Tyr-SRIF-14 and 125I-N-Tyr-SRIF-28 binding sites in the brain is identical. In conclusion, the present results are consistent with a neurotransmitter and neurohormonal role for SRIF-14 and SRIF-28. The function of SRIF-28(1-12) in brain remains to be elucidated. Additionally, a differential role for SRIF-14 and SRIF-28 both in adenohypophysis and brain cannot be ascertained at the present time. 相似文献
7.
M Maletti M Andersson J C Marie G Rosselin V Mutt 《The Journal of biological chemistry》1992,267(22):15620-15625
Somatostatin-28 (SRIF-28) preferring receptors were solubilized from hamster beta cell insulinoma using the zwitterionic detergent 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate. The binding of the iodinated [Leu8-D-TRP22-Tyr25]SRIF-28 analog (referred to as 125I[LWY] SRIF-28) to the solubilized fraction was time-dependent, saturable, and reversible. Scatchard analysis of equilibrium binding data indicated that the solubilized extract contained two classes of SRIF-28-binding sites: a high affinity site (Kd = 0.3 nM and Bmax = 1 pmol/mg protein) and a low affinity site (Kd = 13 nM and Bmax = 4.7 pmol/mg protein). The binding of 125I[LWY]SRIF-28 to solubilized SRIF-28 receptors was sensitive to the GTP analog guanosine-5'-O-thiotriphosphate, suggesting that receptors are functionally linked to a G-protein. By anion-exchange chromatography of the solubilized extract followed by chromatography on wheat germ agglutinin, a 46-fold purification of SRIF-28 receptors was obtained. At this stage of purification, only high affinity sites were found (Kd = 1 nM) and the GTP effect was not maintained. A specific protein of 37 kDa was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after photoaffinity labeling. We suggest that this protein is the putative SRIF-28 receptor or a subunit thereof. 相似文献
8.
Lin X Otto CJ Cardenas R Peter RE 《Canadian journal of physiology and pharmacology》2000,78(12):1053-1066
Somatostatin (SRIF or SS) is a phylogenetically ancient, multigene family of peptides. SRIF-14 is conserved with identical primary structure in species of all classes of vertebrates. The presence of multiple SRIF genes has been demonstrated in a number of fish species and could extend to tetrapods. Three distinct SRIF genes have been identified in goldfish. One of these genes, which encodes [Pro2]SRIF-14, is also present in sturgeon and African lungfish, and is closely associated with amphibian [Pro2,Met13]SRIF-14 gene and mammalian cortistatin gene. The post-translational processing of SRIF precursors could result in multiple forms of mature SRIF peptides, with differential abundance and tissue- or cell type-specific patterns. The main neuroendocrine role of SRIF-14 peptide that has been determined in fish is the inhibition of pituitary growth hormone secretion. The functions of SRIF-14 variant or larger forms of SRIF peptide and the regulation of SRIF gene expression remain to be explored. Type 1 and type 2 SRIF receptors have been identified from goldfish and a type 3 SRIF receptor has been identified from an electric fish. Fish SRIF receptors display considerable homology with mammalian counterparts in terms of primary structure and negative coupling to adenylate cyclase. Although additional types of receptors remain to be determined, identification of the multiple gene family of SRIF peptides and multiple types of SRIF receptors opens a new avenue for the study of physiological roles of SRIF, and the molecular and cellular mechanisms of SRIF action in fish. 相似文献
9.
J. M. Diez G. Giannicot † E. McLeant † E. M. Donaldson †‡ 《Journal of fish biology》1992,40(6):887-893
Porcine galanin, somatostatins (SRIF-25 and SRIF-28) and invariant SRIF-14, known to have inhibitory-stimulatory actions on growth hormone (GH) secretion in higher vertebrates, were tested for their ability to affect plasma GH levels in coho salmon. Peptides were administered by intraperitoneal injection of 10 or 100 ng g−1 body weight. All three SRIFs decreased plasma GH concentrations, their activity following the order SRIF-14 > SRIF-28 > SRIF-25. Galanin and an anti-SRIF produced pronounced, although transient increases in plasma GH. 相似文献
10.
Lin X Peter RE 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2001,129(2-3):543-550
Somatostatin (SRIF) is a multigene family of peptides. SRIF-14 is conserved with identical primary structure in species across the vertebrates. The presence of multiple SRIF genes has been demonstrated in a number of fish species. Notably, three distinct SRIF genes have been identified in goldfish. One of these genes, which encodes [Pro(2)]SRIF-14, has also been identified in sturgeon and African lungfish, and is closely associated with the amphibian [Pro(2),Met(13)]SRIF-14 gene and mammalian cortistatin gene. The main neuroendocrine role of SRIF-14 peptide that has been determined in fish is the inhibition of pituitary growth hormone secretion. The functions of SRIF-14 variant or larger forms of SRIF peptide and the regulation of SRIF gene expression remain to be explored. Type one and two SRIF receptors have been identified from goldfish and type three SRIF receptor from an electric fish. Fish SRIF receptors display considerable homology to mammalian counterparts in terms of primary structure and negative coupling to adenylate cyclase. The identification of the multiple gene family of SRIF peptides and multiple types of SRIF receptors in fish opens a new avenue for the study of physiological roles of SRIF, and the molecular and cellular mechanisms of SRIF actions in fish. 相似文献
11.
Rat cerebral cortex synaptosomes were exposed in superfusion to various depolarizing stimuli and the release of somatostatin-like immunoreactivity (SRIF-LI) was measured by means of a radioimmunoassay procedure. High KCl (9-50 mM) concentration dependently evoked SRIF-LI release; the evoked overflow reached a plateau at 25 mM KCl and was completely abolished when Ca2+ ions were omitted from the superfusion medium, independently of the concentration of KCl used. The 15 mM K(+)-evoked release of SRIF-LI increased sharply as the Ca2+ concentration was raised to 0.8 mM, then leveled off and reached a plateau at 1.2 mM. The 15 mM K(+)-evoked overflow, but not the spontaneous outflow, was partially decreased (50%) by 1 microM tetrodotoxin. The presence in the superfusion fluid of a mixture of peptidase inhibitors did not improve the recovery of SRIF-LI both in the absence and in the presence of high K+. Exposure of synaptosomes to veratrine (1-50 microM) induced release of SRIF-LI in a concentration-dependent way. The effect of the alkaloid was strictly Ca2+ and tetrodotoxin sensitive. Replacement of extracellular Na+ by sucrose caused an acceleration of the spontaneous SRIF-LI outflow that was inversely correlated to the Na+ content in the superfusion medium. The release evoked by the sodium-deprived media did not exhibit any calcium dependence. HPLC analysis of the samples collected during superfusion showed that greater than 90% of the SRIF-LI released either during the spontaneous outflow or by 15 mM KCl was represented by SRIF-14 (SRIF-28(14-28]. These values reflected the ratio SRIF-14/SRIF-28 found in synaptosomes at the end of the experiments. 相似文献
12.
Fingerprint analysis of bovine hypothalamic preprosomatostatin. Identification of somatostatin-28 at the C terminus 总被引:1,自引:0,他引:1
Messenger RNA from bovine hypothalami was used to direct the synthesis in vitro of a precursor to somatostatin (SRIF) of Mr 15,500. Specific antibodies, raised against the chemically synthesized tetradecapeptide SRIF-14, were used for the preliminary characterization. The radioactively labelled preprosomatostatin was then cleaved by trypsin or cyanogen bromide and the products were assayed by two-dimensional fingerprinting techniques. The results conclusively demonstrated the presence of the tetradecapeptide SRIF-14 sequence and its naturally occurring N-terminally extended form, SRIF-28. This 28-amino-acid sequence was shown to occupy the C terminus of the 15,500-dalton precursor and is probably preceded by basic amino acid(s). 相似文献
13.
The two groups of neurosecretory cells producing neuropeptides related to somatostatin (SRIF) and methionine-enkephalin (met-enkephalin), previously high-lighted in the brain of adult migratory locusts, were detected by immunofluorescent techniques during the embryonic development of these insects. The earliest detection of these neurosecretory products occurred firstly in the terminal arborizations, then in the fibres, and finally in the perikarya. SRIF-like material is present in the corpora cardiaca already four days before hatching, i.e. at two-thirds of embryonic life, whereas immunoreactivity can be detected only after hatching in the perikarya located in the pars intercerebralis. The synthesis of met-enkephalin-like neuropeptide starts in the four cells of this system at least two days before hatching as shown by the immunofluorescence in the terminal arborizations along the tractus I to the corpora cardiaca. SRIF-like and met-enkephalin-like neurosecretory products are synthesized and carried to their release areas whilst the formation of brain structures and of the corpora cardiaca has not yet been completed. 相似文献
14.
Senescence marker protein 30 (SMP30) expression in eukaryotic cells: existence of multiple species and membrane localization 总被引:1,自引:0,他引:1
Senescence marker protein (SMP30), also known as regucalcin, is a 34 kDa cytosolic marker protein of aging which plays an important role in intracellular Ca(2+) homeostasis, ascorbic acid biosynthesis, oxidative stress, and detoxification of chemical warfare nerve agents. In our goal to investigate the activity of SMP30 for the detoxification of nerve agents, we have produced a recombinant adenovirus expressing human SMP30 as a fusion protein with a hemaglutinin tag (Ad-SMP30-HA). Ad-SMP30-HA transduced the expression of SMP30-HA and two additional forms of SMP30 with molecular sizes ~28 kDa and 24 kDa in HEK-293A and C3A liver cells in a dose and time-dependent manner. Intravenous administration of Ad-SMP30-HA in mice results in the expression of all the three forms of SMP30 in the liver and diaphragm. LC-MS/MS results confirmed that the lower molecular weight 28 kDa and 24 kDa proteins are related to the 34 kDa SMP30. The 28 kDa and 24 kDa SMP30 forms were also detected in normal rat liver and mice injected with Ad-SMP30-HA suggesting that SMP30 does exist in multiple forms under physiological conditions. Time course experiments in both cell lines suggest that the 28 kDa and 24 kDa SMP30 forms are likely generated from the 34 kDa SMP30. Interestingly, the 28 kDa and 24 kDa SMP30 forms appeared initially in the cytosol and shifted to the particulate fraction. Studies using small molecule inhibitors of proteolytic pathways revealed the potential involvement of β and γ-secretases but not calpains, lysosomal proteases, proteasome and caspases. This is the first report describing the existence of multiple forms of SMP30, their preferential distribution to membranes and their generation through proteolysis possibly mediated by secretase enzymes. 相似文献
15.
Badway AC West FM Tente SM Blake AD 《Biochemical and biophysical research communications》2004,319(4):1222-1227
Somatostatin (somatotropin release inhibitory factor; SRIF) is an endogenous peptide produced at sites of inflammation, making the SRIF a candidate in regulating vascular inflammation. We have used primary human coronary artery endothelial cells (hCAEC) as a model to study SRIF's vascular actions. RT-PCR analysis of hCAEC total mRNA demonstrated the presence of the sst(4) receptor subtype, providing a target for SRIF intracellular signaling. Western blotting with phospho-specific ERK1/2 antibodies showed that SRIF-14 acutely inhibited basal phosphorylation of the extracellular regulated kinases (ERK1/2) by 80%. In addition, SRIF-14 treated hCAEC cell lysates showed a 2.6-fold increase in phosphatase activity, which was inhibited by sodium vanadate. Furthermore, SRIF-14 appeared to be anti-inflammatory in hCAEC as IL-1beta-induced adhesion molecule expression was reduced by 50%. Together, these results show that the coronary artery endothelium is a direct target of SRIF action. 相似文献
16.
Octapeptide analogs of somatostatin exhibiting greatly enhanced in vivo and in vitro inhibition of growth hormone secretion in the rat 总被引:1,自引:0,他引:1
W A Murphy M L Heiman V A Lance I Mezo D H Coy 《Biochemical and biophysical research communications》1985,132(3):922-928
Analogs of a superactive somatostatin (SRIF) octapeptide (code named SMS 201-995 (1)) were synthesized using solid-phase synthetic methodology and assayed for their ability to inhibit growth hormone release from cultured rat anterior pituitary cells and in sodium pentobarbital-anesthetized rats. One analog: (Formula: see text) exhibited greatly enhanced in vitro inhibitory activity (greater than 1,000x) relative to both the parent octapeptide molecule and to the 14 amino acid SRIF molecule. This analog which was also very potent in vivo contains a tyrosine residue and, given its high in vitro activity, may be of investigative importance as a radioiodinated ligand in receptor assays. An octapeptide retro-inverso analog also exhibited significant SRIF-like activity. Several very low activity octapeptide analogs were synthesized and were found to be devoid of SRIF-antagonist activity. A dodecapeptide analog previously shown to be superactive in vivo also demonstrated high in vitro activity. 相似文献
17.
Dr. M. Olivereau F. Ollevier F. Vandesande J. Olivereau 《Cell and tissue research》1984,238(2):289-296
Summary Immunocytochemical investigations show that somatostatin (SRIF)-like immunoreactive material is present in the brain and the pituitary of nine different species of teleosts. In the brain, immunoreactive perikarya and fibers are observed in the preoptic periventricular nucleus, the entopeduncular nucleus, the anterior periventricular nucleus, and the nucleus lateralis tuberis. In the pituitary, SRIF-like-immunoreactive fibers occur in the proximal pars distalis (PPD), which contains the growth hormone (GH)-secreting cells. Nerve fibers are scattered among GH cells (cyprinids), or end on the basal lamina at the neuroglandular interface of the PPD (eel, salmonids). In the eel, the proximal neurohypophysis does not penetrate deeply into the PPD that is very poorly vascularized. In some species, e.g. Myoxocephalus, SRIF-like immunoreactive fibers are also observed in the caudal neurohypophysis, and even among MSH cells of the pars intermedia.In long-term starved carps and eels, the amount of SRIF-like material in the pituitary is clearly reduced. A possible role of SRIF in the concomitant stimulation of GH cells is discussed. 相似文献
18.
The bovine maturation-associated sperm membrane antigen CD52-like molecule has been analysed using a mouse anti-sperm monoclonal antibody developed against bull spermatozoa. The antigen recognised by monoclonal antibody IVA-543 was detected on blood mononuclear cells (including lymphocytes and monocytes) and on a minor population of polymorphonuclear leukocytes. The bovine CD52-like molecule is secreted by the epididymal epithelium and then it is inserted into the sperm membrane during the epididymal transport in the distal part of epididymis. The CD52-like molecule was absent from spermatozoa derived from testes, and the highest proportion of IVA-543-reactive sperm was observed in the cauda epididymis (91.6%).This study has shown that the new molecule identified on bovine cells has properties analogous to those previously described for CD52 molecules in man, mouse, rat, monkey, and dog. 相似文献
19.
Somatostatin (SRIF) is a peptide hormone that is synthesized as part of a larger precursor, prepro-SRIF, consisting of a signal peptide and a proregion of 80-90 amino acids. The mature hormone exists as two different bioactive species. In addition to the most common form, which is a 14-residue peptide, there is also a 14-amino acid NH2-terminally extended form of the tetradecapeptide, SRIF-28. In mammals a single prepro-SRIF molecule undergoes tissue-specific processing to generate the mature hormone, whereas in some species of fish separate genes encode two distinct but homologous precursors, prepro-SRIF-I and -II, that give rise to SRIF-14 and -28, respectively. To investigate the molecular basis for differential processing of the prohormones, we have expressed their cDNAs in heterologous cells. Previously, we demonstrated that prepro-SRIF-I was efficiently and accurately processed in rat pituitary growth hormone (GH3) cells to generate the same hormone as synthesized in pancreatic islet D-cells, namely SRIF-14 (Stoller, T., and Shields, D. (1989) J. Biol. Chem. 264, 6922-6928). We have now compared the proteolytic processing of pro-SRIF-II to that of pro-SRIF-I in these cells. In contrast to pro-SRIF-I, pro-SRIF-II was neither processed nor secreted. Instead, greater than 70% of the precursor was degraded intracellularly in a post-trans Golgi network compartment which was inhibited by weak bases. Brefeldin A treatment prevented degradation, suggesting that turnover of the remaining pro-SRIF-II occurred after exit from the endoplasmic reticulum/intermediate compartment and prior to arrival at the trans Golgi network. The intracellular degradation of the precursor was unexpected, since heterologous cells which do not cleave prohormones generally secrete the unprocessed precursor. We speculate that unique structural domains within each precursor are recognized by the sorting apparatus in GH3 cells, thereby targeting the molecules to different intracellular organelles. 相似文献
20.
The HLA-D region of individuals with the DRw11, w52, DQw3 haplotype encodes multiple molecular products of three distinct subregions, DR, DP, and DQ. Since each molecule can carry multiple stimulatory epitopes, the repertoire of allogeneic T-cell responses to determinants of this haplotype can be quite large. In the present experiments, alloreactive cloned T-cell lines recognized six distinct epitopes associated with DRw11, DRw52, DQw3 haplotypes. Panel studies established that three epitopes were DRwll-like and three were DRw52-like. Blocking with monoclonal antibodies showed that two DRw11-like epitopes were carried by DR-subregion products and one DRwll-like epitope was carried by DQ-subregion molecules. DRw52-like epitopes were detected on separate DR subregion-encoded molecules. One of them carried both DRwl1-and DRw52-like epitopes, the other carried two of the DRw52-like epitopes. These epitopes, which represent functional units that trigger T-cell responses, can be detected at the present time only with the methods used in this report. Conventional allogeneic T-cell responses represent the summation of responses to multiple epitopes encoded by different D-subregion genes. 相似文献