PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “Enkephalin, Leucine”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 19 recordsLinked to original sources

[The effect of methionine enkephalin, leucine enkephalin and the hexapeptide, dalargin on the morphology of cultured neurons].

Cell cultures derived from brain, ganglion trigeminale or retina of 5- and 9-day-old chick embryos and from brain stem or hippocampus of 16-18 day-old fetal rats were treated with Met5-, Leu5-enkephalin, or Dalargin--a synthetic hexapeptide structurally related to Leu5-enkephalin--and evaluated morphologically. The treatment of the nervous tissue cultures with the peptides markedly affected their growth pattern. Aggregates of glial and neuronal cells were surrounded by flat cells forming monolayer network structures. In cultures of brain from 5 day-old chick embryos the proliferation of the cells was significantly stimulated. No growth of neurites was observed. It is discussed that the tested Enkephalins and in the same way the Dalargin may play an important role in the early neuronal development. Their activity in phase 0 and phase 1 of the life history of nerve cells indicates aspects of temporary peptide influences. Excess of glial cells caused by the tested peptides did not enhance neuronal differentiation. The transdifferentiation of developing receptor cells into ependymal cells during the adaptation process in retina cultures as a special effect of Dalargin is discussed. The present findings suggest that the influence of (opioid) peptides in vitro may be detected morphologically and that nervous tissue culture is a valuable tool for differentiating neuronotrophic neurite-promoting and neurotoxic drug effects.

Animals↗

Multi-stage tandem mass spectrometry of metal cationized leucine enkephalin and leucine enkephalin amide.

We have examined the multi-stage collision induced dissociation (CID) of metal cationized leucine enkephalin, leucine enkephalin amide, and the N-acetylated versions of the peptides using ion trap mass spectrometry. In accord with earlier studies, the most prominent species observed during the multi-stage CID of alkali metal cationized leucine enkephalin are the [b(n) + 17 + Cat]+ ions. At higher CID stages (i.e. >MS(4)), however, dissociation of the [b2 + 17 + Cat]+ ion, a cationized dipeptide, results in the production of [a(n) -1 + Cat]+ species. The multi-stage CID of Ag+ cationized leucine enkephalin can be initiated with either the [b(n) -1 + Ag]+ or [b(n) + 17 + Ag]+ ions produced at the MS/MS stage. For the former, sequential CID stages cause, in general, the loss of CO, and then the loss of the imine of the C-terminal amino acid, to reveal the amino acid sequence. Similar to the alkali cationized species, CID of [b2 -1 + Ag]+ produces prominent [a(n) -1 + Ag]+ ions. The multi-stage CID of argentinated peptides is reminiscent of fragmentation observed for protonated peptides, in that a series of (b(n)) and (a(n)) type ions are generated in sequential CID stages. The Ag+ cation is similar to the alkali metals, however, in that the [b(n) + 17 + Ag]+ product is produced at the MS/MS and MS3 stages, and that sequential CID stages cause the elimination of amino acid residues primarily from the C-terminus. We found that N-acetylation of the peptide significantly influenced the fragmentation pathways observed, in particular by promoting the formation of more easily interpreted (in the context of unambiguous sequence determination) dissociation spectra from the [b2 + 17 + Li]+, [b2 + 17 + Na]+ and [b2 -1 + Ag]+ precursor ions. Our results suggest, therefore, that N-acetylation may improve the efficacy of multi-stage CID experiments for C-terminal peptide sequencing in the gas phase. For leucine enkephalin amide, only the multi-stage CID of the argentinated peptide allowed the complete amino acid sequence to be determined from the C-terminal side.

Cations↗

Methionine-enkephalin, leucine-enkephalin methionine-enkephalin-Arg6-Phe7 and methionine-enkephalin-Arg6-Gly7-Leu8 in human pheochromocytoma.

Methionine-enkephalin(met-enkephalin)-, leucine-enkephalin(leu-enkephalin)-, methionine-enkephalin-Arg6-Phe7(met-enkephalin-Arg-Phe)- and methionine-enkephalin-Arg6-Gly7-Leu8(met-enkephalin-Arg-Gly-Leu)-like immunoreactivities(-LI) were studied in 16 pheochromocytomas by radioimmunoassays (RIAs) for these four opioid peptides. Met-enkephalin-Arg-Phe-LI and met-enkephalin-Arg-Gly-Leu-LI existed together with met-enkephalin-LI and leu-enkephalin-LI in 16 pheochromocytomas. Significant positive correlations were observed among contents of these four opioid peptides in 16 pheochromocytomas. The concentrations of these four opioid peptides in epinephrine producing pheochromocytomas were much higher than those in norepinephrine producing tumors. HPLC and gel exclusion chromatography followed by the RIAs showed the presence of met-enkephalin, leu-enkephalin, met-enkephalin-Arg-Phe and met-enkephalin-Arg-Gly-Leu together with their high molecular weight forms. These results indicate the co-existence of met-enkephalin, leu-enkephalin, met-enkephalin-Arg-Phe, met-enkephalin-Arg-Gly-Leu and their high molecular weight forms derived from preproenkephalin A in human pheochromocytomas and suggest the association of preproenkephalin A synthesis with epinephrine production in human pheochromocytomas.

Adrenal Gland Neoplasms↗

Demonstration and characterization of immunoreactive methionine-enkephalin, leucine-enkephalin, methionine-enkephalin-Arg6-Gly7-Leu8 and methionine-enkephalin-Arg6-Phe7 in human phaeochromocytoma.

To elucidate whether or not human phaeochromocytoma contains methionine-enkephalin-Arg6-Gly7-Leu8 (Met-enkephalin-Arg-Gly-Leu) and methionine-enkephalin-Arg6-Phe7 (Met-enkephalin-Arg-Phe) together with methionine-enkephalin (Met-enkephalin) and leucine-enkephalin (Leu-enkephalin), all of which are known to exist in the same precursor molecule (preproenkephalin A), we examined extracts from 16 phaeochromocytomas using high performance liquid chromatography (HPLC) and gel exclusion chromatography coupled with radioimmunoassays (RIAs) for these four opioid peptides. Met-enkephalin-Arg-Gly-Leu-like immunoreactivity (-LI) and Met-enkephalin-Arg-Phe-LI existed together with Met-enkephalin-LI and Leu-enkephalin-LI in 16 phaeochromocytomas. There was a wide variation in contents of Met-enkephalin-LI, Leu-enkephalin-LI, Met-enkephalin-Arg-Gly-Leu-LI and Met-enkephalin-Arg-Phe-LI. Significant positive correlations were observed among the contents of these four opioid peptides in 16 phaeochromocytomas. HPLC and gel exclusion chromatography followed by the RIAs showed the presence of Met-enkephalin, Leu-enkephalin, Met-enkephalin-Arg-Gly-Leu and Met-enkephalin-Arg-Phe together with their high molecular weight forms, which existed in variable amounts. Molar ratios of the contents of these four opioid peptides determined after HPLC varies from case to case. These results indicate the co-existence of Met-enkephalin, Leu-enkephalin, Met-enkephalin-Arg-Gly-Leu and Met-enkephalin-Arg-Phe in human phaeochromocytomas, suggesting the preservation of amino acid sequences of these four opioid peptides even in neoplastic tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

In vitro interactions of opioid peptides with phospholipids. IV. Methionine enkephalin versus leucine enkephalin.

The interaction of methionine and leucine enkephalin with phosphatidylserine and phosphatidylcholine was studied by optical spectroscopy techniques. The data reported indicate that with both peptides the binding is controlled by ionic parameters. They also indicate that the differences in the binding behavior of the two peptides induced by changing these parameters are minor. Non-ionic interactions are also important in the binding phenomena, but the above observations hold in this case as well. Finally, the tridimensional structure of both enkephalins appears to be modified in the presence of phospholipids. Moreover, the changes induced by these lipids appear to differ from one peptide to the other.

Circular Dichroism↗

Parallel distribution of methionine-enkephalin-Arg6-Gly7-Leu8 with methionine-enkephalin, leucine-enkephalin and methionine-enkephalin-Arg6-Phe7 in human and bovine brains.

Using specific radioimmunoassays(RIAs) for methionine-enkephalin(Met-Enk), leucine-enkephalin(Leu-Enk), methionine-enkephalin-Arg6-Gly7-Leu8 (Met-Enk-Arg-Gly-Leu) and methionine-enkephalin-Arg6-Phe7 (Met-Enk-Arg-Phe), we studied the regional distribution of these opioid peptides in human and bovine brains. Met-Enk-Arg-Gly-Leu was distributed in parallel with Met-Enk, Leu-Enk and Met-Enk-Arg-Phe in human and bovine brains. The ratios of molar concentrations of these peptides are almost constant in various regions of human and bovine brains and similar to the ratio of these peptides contained in preproenkephalin A. Gel exclusion chromatography and HPLC coupled with respective RIAs showed the existence of authentic peptides without any detectable high molecular weight forms. These results indicate the parallel distribution of Met-Enk-Arg-Gly-Leu with Met-Enk, Leu-Enk and Met-Enk-Arg-Phe in various regions of human and bovine brains and further suggest that these opioid peptides are derived from the same precursor as that in the adrenal medulla and that the processing of preproenkephalin A is almost complete in human and bovine brains.

Animals↗

Occurrence of methionine-enkephalin-Arg6-Gly7-Leu8 with methionine-enkephalin, leucine-enkephalin and methionine-enkephalin-Arg6-Phe7 in human gastric antrum.

Recent studies on the nucleotide sequence of cloned DNA complementary to mRNA for preproenkephalin from bovine adrenal medulla and human pheochromocytoma have revealed that this precursor molecule contains four copies of methionine-enkephalin (Met-Enk) and one copy each of leucine-enkephalin (Leu-Enk), methionine-enkephalin-Arg6-Gly7-Leu8 (Met-Enk-Arg6-Gly7-Leu8) and methionine-enkephalin-Arg6-Phe7 (Met-Enk-Arg6-Phe7). We have demonstrated the existence of Met-Enk-Arg6-Gly7-Leu8 together with Met-Enk, Leu-Enk and Met-Enk-Arg6-Phe7 in human gastric antrum, using high performance liquid chromatography (HPLC) coupled with radioimmunoassays for these opioid peptides. The ratio of molar concentrations of these peptides in human gastric antrum is similar to the ratio of these peptides contained in preproenkephalin. Furthermore, gel filtration studies on Sephadex G-50 showed that most of immunoreactivities of these peptides were eluted at the elution position of each synthetic peptide without any detectable immunoreactivities at high molecular weight positions. These results indicate the presence of Met-Enk-Arg6-Gly7-Leu8 together with Met-Enk, Leu-Enk and Met-Enk-Arg6-Phe7 in human gastric antrum and further suggest that these opioid peptides are derived from the same preproenkephalin as that in the adrenal medulla and that the processing of preproenkephalin is almost complete in the gut.

Adult↗

Measurement of methionine enkephalin and leucine enkephalin in rat brain regions by high-performance liquid chromatography with coulometric electrochemical detection.

A method is described for the determination of two pentapeptides, methionine enkephalin (H-Tyr-Gly-Gly-Phe-Met-OH) (ME) and leucine enkephalin (H-Tyr-Gly-Gly-Phe-Leu-OH) (LE) in discrete rat brain regions. Separation and quantitation were performed by reversed-phase high-performance liquid chromatography with coulometric detection. Perchloric acid extracts of the tissue after enzyme inactivation by heat treatment were passed through a normal-phase solid-phase extraction diol (COHCOH) column, and endogenous ME and LE were subsequently eluted with methanol. The mobile phase was 1-propanol-phosphate buffer (pH 5.5) (9:91). Eluted samples were detected electrochemically using dual coulometric electrodes operated in screen mode. Each of these enkephalins gave a linear response over the range 40-160 ng/ml cerebellar homogenate (0.8-3.2 ng absolute amount on column). Analytical recoveries of synthetic ME and LE, added to the homogenates, were 70 +/- 3 and 70 +/- 10%, respectively, when compared with enkephalins dissolved in water. The mean between-assay coefficients of variation for synthetic ME and LE were lower than 10.7 and 7.4%, respectively, over the concentration range studied. The within-assay coefficients of variation for synthetic ME and LE were 11.4 and 9.5%, respectively, at the lowest concentration. The present method has been applied to a study determining the levels of endogenous ME and LE in discrete rat brain regions.

Animals↗

The distribution of methionine-enkephalin and leucine-enkephalin in the brain and peripheral tissues.

1 A method is described for the rapid extraction of opioid peptides from the brain and other tissues. The method is based on acid extraction of tissues followed by adsorption of the extract onto Amberlite XAD-2 resin. Elution with methanol separates the enkephalins and alpha-endorphin from beta-endorphin.2 Over 90% of the opioid peptide activity isolated from brain and gut of several species by our method was due to methionine- and leucine-enkephalin. In contrast, the major opioid peptide activity recovered from the pituitary was due to peptides of much greater mol. wt. than the enkephalins.3 An opioid peptide with properties unlike those of the known endorphins or enkephalins was present in brain extracts. This peptide, termed epsilon-endorphin, has an apparent mol. wt. of 700 to 1200; it constituted between 5 to 10% of the total opioid activity in our extracts.4 A differential assay of methionine- and leucine-enkephalin was made either by destroying methionine-enkephalin activity with cyanogen bromide or by separating the peptides by thin layer chromatography.5 The ratio of methionine-enkephalin to leucine-enkephalin varied greatly in different brain regions. The highest proportions of leucine-enkephalin were found in the cerebral cortex and hippocampus.6 Formaldehyde perfusion and fixation of the brain in vivo had no significant effect on the brain content of enkephalin, indicating that proteolytic breakdown is not a major problem in the extraction of these peptides.7 It is suggested that the enkephalins may have a neurotransmitter role in both brain and peripheral tissues and that methionine- and leucine-enkephalin may subserve separate neuronal functions.

Animals↗

Plasma methionine-enkephalin and leucine-enkephalin in normal subjects and patients with pheochromocytoma.

Plasma methionine-enkephalin-like and leucine-enkephalin-like immunoreactivity (met-enkephalin-LI and leu-enkephalin-LI, respectively) in six normal subjects and six patients with pheochromocytoma were determined. The contents of met-enkephalin-LI and leu-enkephalin-LI in two of six pheochromocytomas were 40- to 50-fold higher and those in the other four pheochromocytomas were less than those in normal human adrenal medulla. The former two patients showed high plasma met-enkephalin-LI and leu-enkephalin-LI levels. As plasma catecholamines levels returned to the normal range after extirpation of tumors, met-enkephalin-LI and leu-enkephalin-LI in plasma became undetectable in these patients. In contrast, neither met-enkephalin-LI nor leu-enkephalin-LI was detected in plasma from the latter four patients. Met-enkephalin-LI and leu-enkephalin-LI concentrations were higher in the adrenal vein than in the peripheral vein in three patients. Plasma met-enkephalin-LI and leu-enkephalin-LI increased concomitantly with catecholamines after glucagon stimulation and during a spontaneous attack in a patient with pheochromocytoma. Plasma met-enkephalin-LI changed in parallel with leu-enkephalin-LI in all cases. High performance liquid chromatography coupled with RIAs has shown that met-enkephalin and leu-enkephalin circulate in plasma from a patient with pheochromocytoma as intact pentapeptides. None of normal subjects showed detectable concentrations of met-enkephalin-LI and leu-enkephalin-LI in plasma (more than 5 pg/ml and 3 pg/ml, respectively). It is concluded that met-enkephalin and leu-enkephalin are released concomitantly with catecholamines from pheochromocytomas.

Adrenal Gland Neoplasms↗

[Role of opioid peptide in rheumatoid arthritis--detection of methionine-enkephalin and leucine-enkephalin in synovial tissue].

A relationship between rheumatoid arthritis (RA) and endocrinological abnormality has been suspected for many years. In the present study, we immunohistologically identified the presence of methionine-enkephalin (Met-enk) and leucine-enkephalin (Leu-enk), in synovial tissues collected from RA patients and determined the amount of these peptides and interleukin 1 beta (IL-1 beta) in the culture supernatant of tissue explants by RI and ELISA. Synovial membrane samples obtained by biopsy during artificial joint replacement and plasma samples simultaneously collected from 11 RA patients (11 joint) diagnosed as having classic or definite RA according to the diagnostic criteria of the American Rheumatism Association were used. All 11 patients were females and aged between 36 and 64 years (mean: 45.8 years). As a result of immunohistological standing (ABC method), Met-enk was detected in vascular endothelial cells and Leu-enk in superficial cells, vascular endothelial cells and interstitial cells. Leu-enk concentrations (42.3 +/- 15.8 pg/ml) were higher in the culture supernatant of the synovial membrane than in the plasma (20.7 +/- 13.3 pg/ml). Furthermore, in the culture supernatant the concentration of Leu-enk was positively correlated with that of IL-1 beta (r = 0.789). These findings suggest that Met-enk and Leu-enk are locally produced by synovial tissues and may exert an immunological and/or inflammatory role in RA synovitis.

Adult↗

Methionine-enkephalin and leucine-enkephalin in human sympathoadrenal system and pheochromocytoma.

To elucidate the physiological and pathophysiological significance of methionine- and leucine-enkephalin (Met-and Leu-enkephalin, respectively) in human sympathoadrenal system, the contents of these peptides in normal human sympathetic nervous system, adrenal medulla, and pheochromocytomas were determined by specific radioimmunoassays combined with reverse-phase high-performance liquid chromatography. Met-enkephalin-LI and Leu-enkephalin-LI, respectively) were detected by radioimmunoassay in adrenal glands, adrenal medulla, stellate ganglia, sympathetic trunks, and celiac ganglia, and their contents in adrenal medulla were highest. Existence of authentic Met- and Leu-enkephalin was confirmed by reverse-phase high-performance liquid chromatography. Met-enkephalin was approximately 74% of Met-enkephalin-LI, whereas Leu-enkephalin was approximately 30% of Leu-enkephalin-LI in human adrenal medulla. The ratio of Met- to Leu-enkephalin was 2.6 in human adrenal medulla, whereas it was higher in sympathetic ganglia or trunks. In four cases of pheochromocytoma marked difference in Met- and Leu-enkephalin contents was found between medullary and extramedullary tumors. The contents were about three orders higher and the Met- to Leu-enkephalin ratio was lower in medullary than in extramedullary pheochromocytomas, reflecting the tissues where the tumors arose. These results suggest the physiological roles of Met- and Leu-enkephalin in sympathetic nervous system and adrenal glands and their pathophysiological significances in pheochromocytomas.

Adrenal Gland Neoplasms↗

Methionine-enkephalin and leucine-enkephalin increase interleukin-1 beta release in mixed glia cultures.

Interleukin-1 beta (IL-1 beta) is synthesized in the brain in response to LPS. Excessive IL-1 beta expression is observed in neurodegenerative diseases. The aim of this study was to evaluate the effects of methionine-enkephalin (ME) and leucine-enkephalin (LE) on the baseline and LPS-activated release of IL-1 beta in rat mixed glia cultures. ME and LE increased LPS-induced IL-1 beta release, which was not blocked by naloxone. Both ME and LE increased the baseline release of IL-1 beta, which was completely blocked by naloxone pretreatment. Mixed glia cultures deprived of microglia (by shaking and incubating with L-leucine methyl ester) did not release IL-1 beta, which indicates microglia as a source of the changes in IL-1 beta release. The results of the study suggest that neurons may regulate glial activity through releasing enkephalins.

Animals↗

Immunohistochemical evidence for the coexistence of substance P, thyrotropin-releasing hormone, GABA, methionine-enkephalin, and leucin-enkephalin in the serotonergic neurons of the caudal raphe nuclei: a dual labeling in the rat.

By means of dual immunohistochemical labeling on the same brain section examined with a light microscope, the present study reports the presence with serotonin (5-hydroxytryptamine; 5-HT) of gamma-aminobutyric acid (GABA), substance P (SP), thyrotropin-releasing hormone (TRH), leucin-enkephalin (LEU-enk), or methionine-enkephalin (MET-enk), within the same neuron in the nuclei raphe magnus, raphe obscurus, and raphe pallidus of the rat. On the one hand, peptides or GABA are detected with specific rabbit antibodies by indirect peroxidase labeling using peroxidase-conjugated Fab fragments, and on the other, 5-HT is detected with a rabbit antibody against the BSA-serotonin conjugate by radio-immunocytochemistry using [125I]-labeled protein A. The possible coexistence of TRH and SP in these neurons is also investigated by using peroxidase labeling and radio-immunocytochemical detection, respectively. In the whole caudal raphe nuclei the proportion of each coexisting peptide with 5-HT appears in decreasing order as: TRH greater than SP greater than MET-enk # LEU-enk greater than GABA. In all instances the level of coexistence differs considerably in B1-B2 vs. B3 cell groups. No SP/TRH dually labeled cells have ever been found in any of the serotonergic nuclei of the caudal raphe. Given the evidence that these raphe nuclei project possibly to the spinal cord, these data constitute an anatomical substrate for the several distinct physiological functions presumably subserved by 5-HT in the cord, namely the modulation of nociception, motor, and autonomic functions.

Animals↗

In vitro release of [5-methionine]enkephalin and [5-leucine]-enkephalin from the rat globus pallidus.

Endogenous [5-methionine]enkephalin (Metenkephalin) and [5-leucine]enkephalin (Leu-enkephalin) are released from perfused slices of rat globus pallidus by increased K(+) in a Ca(2+)-dependent manner. Tissue perfused for 40 min contained only 26% of the Met-enkephalin and 44% of the Leu-enkephalin found in the freshly dissected tissue. After perfusion, the mean (+/-SEM) ratio (wt/wt) of Met-enkephalin to Leu-enkephalin was 3.4 +/- 0.2 compared with 5.8 +/- 0.2 in the fresh tissue. The degradation of trace amounts of synthetic [(3)H]enkephalins in the perfusing medium during stimulated release seems to reflect the accelerated degradation of enkephalin released from the tissue: 63% of the Met-enkephalin and 23% of the Leu-enkephalin were degraded in a medium containing bacitracin (30 mug/ml). The mean ratio (wt/wt) of the Met-enkephalin to the Leu-enkephalin recovered after release by exposure of slices to 50 mM K(+) was 2.7 +/- 0.3. When perfusates were corrected for degradation, this ratio increased to about 5.5 which is higher than that found in the perfused tissue. The differences in release, tissue loss, and catabolism of the two enkephalins may be reflecting differences in the metabolic systems operating on the pentapeptides, but this interpretation will have to be validated by in vivo release experiments. In any event these observations strongly suggest that both enkephalins can be considered candidate neurotransmitters in the rat globus pallidus.

Animals↗