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Biomedical subjects

C H Li

Publications and source records attributed to C H Li.

At least 91 records · Page 5Linked to original sources
PubMed1988

Primary structure of equine pituitary prolactin.

Equine prolactin was determined to be a single chain protein of 199 amino acid containing two tryptophan and six cysteine residues, as found in other mammalian prolactins. The primary sequence of equine prolactin was obtained by automated Edman analyses of S-carboxymethylated protein and proteolytic fragments of modified protein. Of the known prolactin sequences, equine prolactin shows closest homology with porcine (93%) and fin whale (87-91%) prolactins. Genetic mutations have produced changes in 17 of 199 residues of equine prolactin relative to its putative ancestral precursor. Since equine growth hormone has undergone alterations in 4 of 191 residues relative to this putative precursor protein, these results support the theory that prolactins are evolving at a faster rate than growth hormones. Consistent with the previously determined circular dichroic spectrum of equine prolactin, 60% of the protein is predicted to form alpha helices.

Amino Acid Sequence↗
PubMed1988

Human follicular gonadotropin releasing peptide analogs. Evaluation of biological (in vitro) and immunological activity.

Human follicular gonadotropin releasing peptide (hF-GRP) has been shown to stimulate pituitary LH and FSH secretion in vitro. Six hF-GRP analogs have been synthesized and evaluated for gonadotropin releasing activity in a rat anterior pituitary primary cell culture system. A tyrosine analog of hF-GRP, [Tyr4]-hF-GRP, retained comparable biological activity in releasing gonadotropins. However, acetylation of hF-GRP in Ac-hF-GRP greatly reduced the in vitro activity. The shorter segments of hF-GRP, hF-GRP-(5-14), and hF-GRP-(10-14), were tested for LH and FSH releasing activity, and it was found that the decapeptide retained moderate activity while the activity of the pentapeptide was markedly lower than hF-GRP. The baboon alpha 1 antitrypsin-(27-40) peptide, b-alpha 1 AT-(27-40), is relatively less potent in releasing LH than hF-GRP. Interestingly, the baboon peptide is more potent (2.5-fold) in releasing FSH under identical conditions. The effect of hF-GRP in releasing LH and FSH was not affected by the presence of LHRH antagonists in cell culture systems. When these peptides were tested for immunological activity in a hF-GRP radioimmunoassay, it was found that hF-GRP and [Tyr4]-hF-GRP have comparable activities. The C-terminal decapeptide of hF-GRP is more active (1.5-fold) in the RIA, and the C-terminal pentapeptide had only one third of the immunoreactivity. The b-alpha 1-AT-(27-40) failed to cross-react in the RIA even at a concentration of 20 micrograms per tube.

Amino Acid Sequence↗
PubMed1988

Isolation and primary structures of elephant adrenocorticotropin and beta-lipotropin.

Adrenocorticotropin and beta-lipotropin have been isolated and purified from elephant pituitary glands. The primary structures of these two hormones were determined by amino acid and sequence analyses of enzymatically cleaved peptides from the hormones. Peptide purification involved the use of gel filtration, reverse phase high performance liquid chromatography, and paper electrophoresis.

Adrenocorticotropic Hormone↗
PubMed1988

Insulin-like growth factors and the multiplication of Tera-2, a human teratoma-derived cell line.

A human teratoma cell line (Tera-2) was grown in serum-free medium, and the population multiplication was stimulated by the addition of somatomedins/insulin-like growth factors (IGFs). Both IGF-I and IGF-II gave maximal stimulation when added daily at 10 ng ml-1. The IGFs did not substantially change the labelling index of the cells, and the IGFs appeared to exert their effect on population multiplication by increasing cell survival. Membranes isolated from Tera-2 cells displayed both type 1 and type 2 IGF receptors.

Cell Division↗
PubMed1988

Insulin-like growth factor-I stimulates oxytocin and progesterone production by bovine granulosa cells in culture.

The effect of insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF), fibroblast growth factor (FGF) and nerve growth factor (NGF) on production of oxytocin and progesterone by cultured bovine granulosa and luteal cells was studied. Secretion of oxytocin was stimulated, in a dose-dependent manner, by IGF-I at 48 and 120 h of culture to levels much higher than those after stimulation with LH, FSH, EGF, FGF or NGF. A similar effect of IGF-I was observed for progesterone but, in contrast to oxytocin, secretion of progesterone was not increased by EGF, NGF or FGF. During primary culture, for 4 h, of dispersed bovine luteal cells obtained from corpora lutea between days 4 and 10 of the oestrous cycle, all the growth factors tested failed to stimulate secretion of oxytocin or progesterone. The data suggest the relevance of growth factors (especially IGF-I) for ovarian physiology and their possible importance for differentiation of follicles and luteinization.

Animals↗
PubMed1988

Beta-endorphin. Receptor binding and peripheral opioid activities of [Gln8]-, [Trp27]-, and [Tyr31]-analogs.

Three beta h-EP analogs which show different extents of alteration in analgesic potency by substitution of a single amino acid residue were assayed for their peripheral opioid activity and the binding to opioid mu-receptor to determine the relationships among the opioid activities obtained from different assays. In the guinea pig ileum assay, [Gln8]-beta h-EP showed a higher inhibitory activity than the parent peptide, [Tyr31]-analog had the same potency as beta h-EP, while [Trp27]-analog retained only one fourth the potency of beta h-EP. Assayed on the vas deferens of the mouse and the rat, all three substituted beta h-EP analogs exhibited a lower potency than their parent peptide. Receptor binding assay using [3H]-dihydromorphine as the primary ligand showed that [Gln8]-analog had a binding potency 1.5-fold that of beta h-EP, while the potencies of [Tyr31]- and [Trp27]-analogs were not significantly different from that of the parent peptide. No correlation in relative potency was found between vas deferens assays and their mu-receptor binding or analgesic activity. However, the relative potencies of binding to mu-receptor in [Gln8]- and [Tyr31]-analogs were found to be consistent with those of analgesic and guinea pig ileum assays, whereas the binding to beta-EP receptor of all analogs appeared to be related to the charge properties of beta-EP molecule.

Animals↗
PubMed1987

IGF-2 stimulated growth mediated by the somatomedin type 2 receptor.

Human Insulinlike Growth Factor 2 (IGF-2) can promote cell proliferation via the type 2 receptor in K562 cells, a human erythroleukemia cell line with IGF-2/type 2 receptors and insulin receptors but lacking IGF-1/type 1 receptors. Cells are grown in semi-solid agar in the absence and presence of increasing amounts of insulin, IGF-1 and IGF-2. Two strains of K562 cells have been studied, with different concentrations of insulin and IGF-2 receptors. The effect of IGF-2 is proportional to the IGF-2 receptor concentration.

Cell Division↗
PubMed1987

The specificity of the human IGF-2 receptor.

The specificity of the type 2 insulinlike growth factor (IGF) receptor is evaluated in human placenta membranes and the human cell line K562. K562 cells have type 2 but not type 1 IGF receptors. Native IGF-2 isolated from human plasma and synthetic IGF-2 were equipotent in competing with labeled IGF-2 in both systems. Pure IGF-1 isolated from plasma, synthetic IGF-1 and recombinant IGF-1 could not crossreact with the type 2 IGF receptor in concentrations up to 1 microgram/ml in both systems. Studies on placenta membrane were done in the presence of 300 ug/ml insulin to block the type 1 IGF receptors. It is concluded that IGF-1, as well as insulin, cannot crossreact with the human type 2 IGF receptor.

Binding, Competitive↗
PubMed1987

Secondary structure determination of human beta-endorphin by 1H NMR spectroscopy.

The 1H NMR spectra of human beta-endorphin indicate that the peptide exists in random-coil form in aqueous solution but becomes helical in mixed solvent. Thermal denaturation NMR experiments show that in water there is no transition between 24 and 75 degrees C, while a slow noncooperative thermal unfolding is observed in a 60% methanol-40% water mixed solvent in the same temperature range. These findings are consistent with circular dichroism studies by other workers concluding that beta-endorphin is a random coil in water but that it forms 50% alpha-helix or more in mixed solvents. The peptide in the mixed water-methanol solvent was further studied by correlated spectroscopy (COSY) and nuclear Overhauser effect spectroscopy (NOESY) experiments. These allow a complete set of assignments to be made and establish two distinct stretches over which the solvent induces formation of alpha-helices: the first occurs between Tyr-1 and Thr-12 and the second between Leu-14 and extending to Lys-28. There is evidence that the latter is capped by a turn occurring between Lys-28 and Glu-31. These helices form at the enkephalin receptor binding site, which is at the amino terminus, and at the morphine receptor binding site, located at the carboxyl terminus [Li, C. H. (1982) Cell (Cambridge, Mass.) 31, 504-505]. Our findings suggest that these two receptors may specifically recognize alpha-helices.

Amino Acid Sequence↗
PubMed1987

Localization of alpha-inhibin-31 in rodent testis and brain by immunocytochemical procedure and Western blot analysis.

Using an anti-serum against synthetic alpha-inhibin-31, we have studied its cellular locations in rodent testis and brain. Western blot analysis detected several molecular forms of the protein(s). A protein of 17 KDa in size was detected in testis of one month old mice. Two major bands of molecular weights 39 kDA and 21 kDA were detected in mature mouse testis. The 39 kDA protein was also present in male brain, female brain and ovary. No protein similar in size to alpha-inhibin-31 was detected with this antiserum, suggesting that the antibody recognized only the precursor of the alpha-IB-31 molecule. Immunocytochemical studies revealed that these molecules were located in the cytoplasm of the interstitial Leydig cells. The immunocytochemical reaction can be eliminated by pre-incubation with synthetic alpha-inhibin-31. Immunoreactive substances were also demonstrated at the blood-brain barrier on the external surface of the ependymal cells in the third ventricle of the rat brain. In the pituitary, similar pattern of localization was also observed in the cuboidal-columnar epithelial cells that lined the par tuberalis between the anterior and intermediate lobes. The immunoreaction to the neural cells cannot be completely abolished by competition with the synthetic peptide. These observations suggested that alpha-inhibins are located in the Leydig cells of the testis as well as in the brain, pituitary and ovary.

Animals↗
PubMed1987

Stimulation of insulin secretion by beta-endorphins (1-27 & 1-31).

Synthetic human beta-endorphin potentiates insulin secretion by the isolated perfused rat pancreas when glucose is present in the perfusate at concentrations of either 125 or 200 mg/dl, whereas it fails to exert any effect on insulin secretion in the presence of a substimulatory concentration of 100 mg/dl. Similar potentiation of insulin secretion occurred in response to the 1-27 fragment (beta-endorphin1-27) of beta-endorphin. This transient potentiation lasts only 3 to 4 minutes, whereupon secretion returns toward control levels. Thus beta-endorphin produces only a transient spike-like secretory profile similar to the first phase of glucose-induced insulin secretion and it fails to produce any chronic insulin secretory response comparable to the second phase of insulin secretion. The insulinotropic effect of beta-endorphins occurred at concentrations varying from 0.1 to 5.0 ug/ml. These data suggest that beta-endorphin and beta-endorphin1-27 potentiate insulin secretion via a common beta cell opioid receptor, and that beta-endorphin may exert a paracrine control of insulin secretion. However, any such regulation appears to be via short-term alterations in the secretory process per se.

Animals↗
PubMed1987

Heterogeneity of insulin-like growth factor-I affinity for the insulin-like growth factor-II receptor: comparison of natural, synthetic and recombinant DNA-derived insulin-like growth factor-I.

Although insulin-like growth factors (IGF) I and II bind with high affinity to structurally discrete receptors, they bind with a lesser affinity to each other's receptor. We have evaluated the affinity of five different IGF-I preparations (three natural IGF-I preparations, one synthetic preparation, and one recombinant DNA-derived) for the IGF-II receptor in rat placental membranes, 18-54,SF cells and BRL-3A cells. In all tissues tested, the natural IGF-I preparations demonstrated an affinity for the IGF-II receptor which was 10-20% that of IGF-II. However, the recombinant and synthetic IGF-I preparations exhibited substantially lower affinities than natural IGF-I for this receptor, with only 10-25% reduction in (125-I)iodo IGF-II binding at peptide concentrations up to 400 ng/ml. Radioimmunoassay of the natural IGF-I preparations with an antibody directed against the unique C-peptide region of IGF-II demonstrated that contamination of IGF-I preparations with immunoreactive IGF-II could not exceed 5%. These results demonstrate that IGF-I purified from human plasma has a different affinity for the IGF-II receptor than does synthetic or recombinant IGF-I. Furthermore, these data are consistent with the hypothesis that IGF-I, itself, may be heterogeneous, and that subforms may vary in their affinities for the IGF receptors. Alternatively, IGF-I preparations which have been considered to be pure may be contaminated with small amounts of IGF-II, resulting in overestimation of the affinity of IGF-I for the type II IGF receptor.

Adult↗
PubMed1987

Specificity of the glutathione S-transferases in the conversion of leukotriene A4 to leukotriene C4.

We have synthesized the 5,6-LTA4, 8,9-LTA4, and 14,15-LTA4 as methyl esters by an improved biomimetic method with yields as high as 70-80%. We have investigated the catalytic efficiency of the purified cytosolic glutathione S-transferase (GST) isozymes from rat liver in the conversion of these leukotriene epoxides to their corresponding LTC4 methyl esters. Among various rat liver GST isozymes, the anionic isozyme, a homodimer of Yb subunit, exhibited the highest specific activity. In general, the isozymes containing the Yb subunit showed better activity than the isozymes containing the Ya and/or Yc subunits. Interestingly, all three different LTA4 methyl esters gave comparable specific activities with a given GST isozyme indicating that regiospecificity of GSTs was not the factor in determining their ability to catalyze this reaction. Surprisingly, purified GSTs from sheep lung and seminal vesicles showed little activity toward these leukotriene epoxides, indicating a lack of the counterpart of rat liver anionic GST isozyme in these tissues.

Animals↗