Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P01189 (beta-endorphin)
 21,003 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A soluble somatostatin binding factor was detected in cell-free extracts from chicken pancreas. For binding measurements Tyr1-somatostatin was radio-labeled with 125I by the lactoperoxidase technique. Specific radioactivity of about 18.5 MBq/nmol was achieved. Maximal total binding is approximately 0.17 (B/T) in the presence of 30 mg/l pancreatic protein. The specific binding is 0.10 and is suppressed by addition of 1 mg/l synthetic cold cyclic somatostatin. The dose-response curve of synthetic cyclic somatostatin is in the range of 0.6-600 nmol/l. Ca2+ and reduced thiol-reagents inhibit the specific binding. Insulin, glucagon and corticotropin show a low, and luliberin and reduced somatostatin a high cross-reactivity. Molecular weight was estimated by gel filtration and the specific binding molecule was eluted at a Kav = 0.2 on an Ultrogel (AcA 54) column. This corresponds to Mr 40 000. Electrophoretic properties of the binding complex and semipurification by polyacrylamide disc gel electrophoresis: relative mobility of the 125I-Tyr-somatostatin binding complex is about 0.6. Relative mobilities of binding-protein fractions are 0.71 and 0.74. Highest relative specific binding was detected in the (100 000 g) cytosol fractions. Binding with cell-free extracts from the splenic lobe area was 4-fold higher than that from other parts of the chicken pancreas.
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PMID:Somatostatin binding factor from chicken pancreas. 611 81

Iodinated native bovine parathyroid hormone (bPTH(1-84)) was separated from uniodinated hormone by reversed-phase liquid chromatography techniques after lactoperoxidase labeling. Analysis of iodinated residues after enzymatic digestion indicated that the major labeled product was largely monoiodinated on the sole tyrosine residue. This material retained full bioactivity in an in vitro renal adenylate cyclase assay. Binding of 125I-bPTH(1-84) to rabbit renal membranes at 4 degrees C was proportional to membrane protein concentration and was saturable and dissociable. Radioligand binding was inhibited by concentrations of unlabeled bPTH(1-84) required to stimulate adenylate cyclase in the same membrane preparation but was not inhibited by non-PTH peptides other than adrenocorticotropin at high concentrations (greater than 10 microM). Synthetic NH2-terminal analogues of bPTH(1-84) all elicited approximately equivalent inhibition of radioligand binding which was, however, less potent than unlabeled bPTH(1-84), suggesting a role for the carboxyl region of the molecule in the interaction of bPTH(1-84) with its receptor. Activity of the NH2-terminal agonists was similar to bPTH(1-84) in stimulating adenylate cyclase. Although substitution in sequence position one, of serine in human PTH(1-34) for alanine in bPTH(1-34), reduced activity in the adenylate cyclase assay, inhibition of 125I-bPTH(1-84) binding by both peptides and by an analogue of bPTH(3-34) was equivalent, consistent with a minimal contribution of the first 2 residues for receptor binding of the NH2-terminal region of PTH. The results illustrate the utility of the radiolabeled preparation of native bPTH we have developed and emphasize the importance of probing the PTH receptor with an intact hormone to maximize information concerning the mechanism of PTH action.
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PMID:Characterization of the rabbit renal receptor for native parathyroid hormone employing a radioligand purified by reversed-phase liquid chromatography. 629 18

We have studied the behaviour of 125I-labelled alpha-MSH under different experimental conditions. Until now, the chloramine T method had been used by most investigators with variable results. We have tested three other labelling techniques based on 125I mild oxidation: (1) an enzymatic method with lactoperoxidase, (2) a sparingly soluble chloramine method (T.D.G.U.) and (3) modified chloramine T procedure, 'the iodine volatilization method'. Labelled hormone obtained after each kind of iodination was assayed for immunoreactivity. In addition, time course degradation was measured by classical RIA incubation procedures. Charcoal-dextran was used to separate bound and free antigen. We have found chloramine T-iodinated alpha-MSH to be significantly more damaged than preparations obtained by other methods and to be less stable when stored at -18 degrees C. No differences were found between the differently labelled 125I-labelled alpha-MSH fresh preparations in binding to surface receptors of human melanoma cell lines in culture.
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PMID:Comparison and evaluation of different methods for alpha-MSH labelling. 675 23

An analogue of human melanin-concentrating hormone (MCH) suitable for radioiodination was designed in which Tyr13 and Val19 of the natural peptide were replaced by phenylalanyl and tyrosyl residues: [Phe13, Tyr19]-MCH. The peptide was synthesized by the continuous-flow solid-phase methodology using Fmoc-strategy and polyhipe PA 500 and PEG-PS resins. The linear MCH peptides with either acetamidomethyl-protected or free cysteinyl residues were purified to homogeneity and cyclized by iodine oxidation, yielding the final product with the correct molecular weight of 2434.61. Radioiodination of the C-terminal tyrosine was carried out enzymatically using solid-phase bound glucose oxidase/lactoperoxidase, followed by purification on a reversed-phase mini-column and by high-pressure liquid chromatography. The resulting [125I]-[Phe13, Tyr19]-MCH tracer was the first radiolabelled MCH peptide suitable for radioreceptor assay: saturation binding analysis using mouse G4F-7 melanoma cells demonstrated the presence of 1090 MCH receptors per cell. The dissociation constant (KD) was 1.18 x 10(-10) M, indicating high-affinity MCH receptors on these cells. MCH receptors were also found in other cell lines such as mouse B16-F1 and G4F and human RE melanoma cells as well as in PC12 and COS-7 cells. Competition binding analyses with a number of other peptides such as alpha-MSH, neuropeptide Y, substance P and pituitary adenylate cyclase activating peptide, demonstrated that the binding to the MCH receptor is specific. Atrial natriuretic factor was found to be a weak competitor of MCH, indicating topological similarities between MCH and ANF when interacting with MCH receptors.
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PMID:Synthesis and iodination of human (phenylalanine 13, tyrosine 19) melanin-concentrating hormone for radioreceptor assay. 922 84

A photoreactive analogue of human melanin-concentrating hormone was designed, [D-Bpa13,Tyr19-MCH, containing the D-enantiomer of photolabile p-benzoylphenylalanine (Bpa) in position 13 and tyrosine for radioiodination in position 19. The linear peptide was synthesized by the continuous-flow solid-phase methodology using Fmoc-strategy and PEG-PS resins, purified to homogeneity and cyclized by iodine oxidation. Radioiodination of [D-Bpa13,Tyr19]-MCH at its Tyr19 residue was carried out enzymatically using solid-phase bound glucose oxidase/lactoperoxidase, followed by purification on a reversed-phase mini-column and HPLC. Saturation binding analysis of [125I]-[D-Bpa13,Tyr19]-MCH with G4F-7 mouse melanoma cells gave a K(D) of 2.2+/-0.2 x 10(-10) mol/l and a B(max) of 1047+/-50 receptors/cell. Competition binding analysis showed that MCH and rANF(1-28) displace [125I]-[D-Bpa13,Tyr19]-MCH from the MCH binding sites on G4F-7 cells whereas alpha-MSH has no effect. Receptor crosslinking by UV-irradiation of G4F-7 cells in the presence of [125I]-[D-Bpa13,Tyr19]-MCH followed by SDS-polyacrylamide gel electrophoresis and autoradiography yielded a band of 45-50 kDa. Identical crosslinked bands were also detected in B16-F1 and G4F mouse melanoma cells, in RE and D10 human melanoma cells as well as in COS-7 cells. Weak staining was found in rat PC12 phaeochromocytoma and Chinese hamster ovary cells. No crosslinking was detected in human MP fibroblasts. These data demonstrate that [125I]-[D-Bpa13,Tyr19]-MCH is a versatile photocrosslinking analogue of MCH suitable to identify MCH receptors in different cells and tissues; the MCH receptor in these cells appears to have the size of a G protein-coupled receptor, most likely with a varying degree of glycosylation.
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PMID:(D-(p-benzoylphenylalanine)13, tyrosine19)-melanin-concentrating hormone, a potent analogue for MCH receptor crosslinking. 1036 6

A highly specific radioimmunoassay was developed for N-terminal peptide of salmonid proopiocortin using a guinea pig antiserum to the chum salmon peptide (sNPP 1). Since sNPP I has no tyrosine residue nor free N-terminal amino group, a mixture of minor components of sNPP 1, which have extensions of H-Val-LysGly- and H-Lys-Gly- at the N-terminus, were iodinated by the lactoperoxidase method after incorporation of 3-(phydroxyphenyl)-propionate to the terminal amino groups. Plasma and pituitary extracts of several salmonid species showed parallel displacement to the standard hormone. Samples from carp, goldfish, tilapia, and eel, as well as the plasma of hypophysectomized rainbow trout, showed no crossreactivity. Proopiocortin-related hormones isolated from the chum salmon pituitary, including melanotropins, endorphins, corticotropin-like intermediate lobe peptides, and gonadotropin and prolactin showed negligible cross-reactivity. NPP contents in the pars intermedia of rainbow trout and chum salmon were 10 to 15 times greater than those in the pars distalis. Plasma levels of NPP in the mature chum salmon caught in the bay were about 50ng/ml. Plasma NPP levels in the mature chum salmon of both sexes decreased after transfer from seawater to fresh water. Plasma cortisol showed a concomitant change with NPP, thus supporting previous findings that NPP modulates corticotropin action on the trout interrenal.
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PMID:A radioimmunoassay for N-terminal peptide of chum salmon proopiocortin. 2423 34