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Biomedical subjects

Y Shimohigashi

Publications and source records attributed to Y Shimohigashi.

At least 109 records · Page 6Linked to original sources
PubMed1984

Importance of the stereo-orientation of aromatic groups in enkephalins to opiate receptor recognition.

PO +Dehydrophenylalanine (delta Phe) having the E-configuration (delta EPhe ; phenyl and C = O cis) was incorporated into [Leu5]-enkephalin in order to restrict its conformation. Compared with the Z-isomer, in the radio-ligand receptor binding assays, [D-Ala2, delta EPhe4 , Leu5] enkephalin showed drastically decreased potency for the delta and mu opiate receptors, i.e., 260- and 150-fold loss of affinity, respectively. The results strongly indicate that the opiate receptors require the Z-configuration (phenyl and C = O, trans) of the delta Phe4 residue and may require a specific interrelationship between the aromatic rings of the Tyr1 and Phe4 residues in the molecule for binding. The conformation of [Leu5]-enkephalin specific for the delta receptors was analyzed and a comparison made with its crystal structure recently elucidated.

Animals↗
PubMed1984

Role of the carbohydrate moiety of human choriogonadotropin in its thyrotropic activity.

To investigate the role of the carbohydrate moiety of human choriogonadotropin (hCG) in its thyrotropic activity, highly purified hCG and its desialylated subunits were treated with anhydrous HF/anisole (1 h, 0 degree C). The deglycosylated alpha- and beta-subunits were recombined with their native complementary subunits, and the interactions of these hCG congeners with the thyrotropin (TSH) receptor-adenylate cyclase system were investigated using human thyroid membranes. Deglycosylated hCG (dghCG) bound to the high affinity-low capacity TSH-binding sites of thyroid membranes; its equilibrium dissociation constant was lower than that of asialo-hCG (ashCG) (ED50: 2.6 and 6 microM, respectively). Like ashCG, dghCG did not stimulate thyroidal adenylate cyclase, but rather inhibited TSH stimulation of this enzyme in a dose-dependent manner. Thus, dghCG behaved as an antagonist and exhibited an inhibition constant of 0.78 microM while ashCG exhibited a constant of 1.50 microM. As might be predicted from the behavior of dghCG, absence of carbohydrate from either subunit enhanced the ability of the hCG hybrid recombinants to interact with the TSH receptor-adenylate cyclase system. However, only the hybrid recombinant lacking carbohydrate on its alpha-subunit lacked intrinsic thyrotropic activity; the hybrid recombinant lacking carbohydrate on its beta-subunit not only displayed intrinsic thyrotropic activity, but was of even higher potency than intact hCG in stimulating thyroidal adenylate cyclase. These results demonstrate that the carbohydrate moieties of both hCG subunits impede the process of recognition of hCG by the TSH receptor, while the carbohydrate moiety of the alpha-subunit, but not that of the beta-subunit, is essential for the process of hCG activation of thyroidal adenylate cyclase.

Adenylyl Cyclases↗
PubMed1983

Gel high-performance liquid chromatographic studies on the elution behavior of chemically deglycosylated human chorionic gonadotropin and its subunits.

The elution behavior of human chorionic gonadotropin and its subunits, and their desialylated and deglycosylated derivatives was studied on gel high-performance liquid chromatography. Using three TSK G-SW columns, excellent separations were achieved for eleven human chorionic gonadotropin derivatives. In a comparison of the native (30% carbohydrate content), asialo (24%) and HF-deglycosylated (8%) subunits, the estimated values of molecular weights on the basis of a series of reference globular proteins deviated in various degrees from the actual molecular weight. The extent of deviation depended on the carbohydrate content and possibly the location of carbohydrate chains. The data from recombination studies suggest that the beta-subunit is the dominant determinant for their expanded molecular size.

Chorionic Gonadotropin↗
PubMed1983

Differential association of spinal mu, delta and kappa opioid receptors with cutaneous thermal and visceral chemical nociceptive stimuli in the rat.

The intrathecal administration of several mu (morphine), delta (d-ala2-d-leu5-enkephalin, dimeric leu-enkephalin) and mixed mu/delta (beta-endorphin) agonists produced a dose-dependent inhibition of all cutaneous thermal (Tail Flick/Hot Plate) nociceptive responses in the rat. The kappa agonist U50488H had no analgesic potency in thermal nociceptive tests. On a visceral chemical test (writhing) and agonists exerted a powerful suppression of the response. In contrast at doses 10 to 50 times the ED50 on cutaneous thermal tests, delta agonists had no effect on the writhing response. At higher intrathecal doses, delta ligands produced flaccidity. These observations suggest the existence of three discriminable populations of opioid receptors in the spinal cord whose activation has different effects on the animals response to noxious stimuli.

Animals↗
PubMed1983

Synthesis and biological activity of delta Phe4-enkephalins.

The delta Phe4-enkephalins have been synthesized and examined in an in vitro receptor binding assay and an in vivo tail flick analgesia test. The delta Phe4 residue was derived from Boc-Gly-Phe(beta-OH)-OH by spontaneous dehydration and azlactonization. The dipeptide azlactone was coupled directly with H-Leu-OBzl to yield a tripeptide which was converted into the pentapeptides after stepwise coupling with two amino acids using the water soluble EDC-HOBt method. Dehydroenkephalins were liberated with hydrogen fluoride in the presence of anisole. In the radioligand binding assay which did not contain an enzyme inhibitor [D-Ala2, delta Phe4, Leu5] enkephalin was almost twice as active as saturated [D-Ala2, D-Leu5]-enkephalin. The delta Phe4-enkephalins exhibited a considerably diminished activity as compared with the saturated peptide in the in vivo analgesic assay. These results are discussed with regard to the enzyme stability and receptor preference of dehydroenkephalins.

Analgesia↗
PubMed1983

Dehydro-enkephalins. VIII. Attempted synthesis of dehydrotyrosine-enkephalins.

Two kinds of dehydropeptide analogs of enkephalin containing a delta Tyr unit at the N-terminus have been synthesized by coupling Boc-delta Tyr-(Cl2 Bzl)-OH with amino acid amides and tetrapeptide esters using the water soluble carbodiimide-HOBt method. Pentapeptides consisting of delta Tyr1, and delta Phe4 or delta Leu5 were also prepared. Ultraviolet difference spectroscopy was important in the characterization of the dehydro moieties, delta Tyr, delta Phe and delta Leu. Attempts to liberate delta Tyr1-enkephalins have been unsuccessful because of the instability of an N-terminal delta Tyr residue having p-phenolic group in the side chain.

Enkephalins↗
PubMed1982

Dimeric pentapeptide enkephalin: a novel probe of delta opiate receptors.

A dimeric pentapeptide enkephalin (DPE2) consisting of two molecules of [D-Ala 2, Leu 5] enkephalin linked at C-terminal leucine with ethylenediamine, (H-Tyr-D-Ala-Gly-Phe-Leu-NH-Ch2)2 is a bivalent ligand for the delta enkephalin receptors of rat brain and neuroblastoma-glioma hybrid (NG108-15) cells. This new enkephalin analog shows dramatically increased affinity in radioligand assays using whole brain membranes when delta but not mu specific radioligands are employed. When membranes from NG108-15 cells are used, the dimer shows greatly increased activity irrespective of the mu or delta specificity of the tracer. The dimer DPE2 shows a four-fold, "sodium shift" in its IC50 for competition with [3H]naloxone, suggestive of agonist behavior. Agonist activity was confirmed by demonstrating that DPE2 inhibits cyclic AMP production in prostaglandin E1 stimulated NG108-15 cells, and by demonstrating very high potency in the mouse vas deferens bioassay. DPE2 binds to the same delta sites as the delta-selective monomer [D-Ala2, D-Leu5] enkephalin, since the two ligands show complete crossdisplacement. Radiolabeled 3H-DPE2 shows a five-fold higher affinity constant, a 2.5-fold higher association rate constant, and a two-fold lower dissociation rate than the monomer. These results are consistent with the hypothesis that the dimeric pentapeptide enkephalin can bridge two delta receptors. This enkephalin dimer provides a valuable new probe of opiate receptors and their organization in cell membranes.

Animals↗
PubMed1982

Dehydro-enkephalins. III. Synthesis and biological activity of [delta Ala2, Leu5]-enkephalin.

[delta Ala2, Leu5]-enkephalin has been prepared and shown to be more active than the parent saturated enkephalin in a binding assay using rat brain membranes and [3H]dihydromorphine as a tracer. In a comparison of potencies against [3H]dihydromorphine and [3H]-[D-Ala2, D-Leu5]-enkephalin as tracers, [delta Ala2, Leu5]-enkephalin showed preference for micro opiate receptors, possibly due to the hydrophobicity of the delta Ala2 residue. A synthetic tetrapeptide enkephalin [delta Ala2]-desLeu5-enkephalin had weak activity and high selectivity for the micro receptors. O-Acylation of a serine residue in the peptide was achieved by coupling between the peptide and a carboxylic acid using DCC and a catalytic amount of 4-dimethylaminopyridine.

Animals↗