Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The human progesterone receptor A and B isoforms (hPR-A and hPR-B) were stably transfected in Chinese Hamster Ovary (CHO) cells in the presence or absence of the mouse mamma tumor virus (MMTV) promoter and luciferase (LUC) reporter gene. In this way four stably transfected CHO cell lines, i.e. hPR-A, hPR-B, hPR-A-MMTV-LUC and hPR-B-MMTV-LUC cells, were prepared. hPR-A and -B isoforms were compared by binding and transactivation analysis for 14 progestagens and 7 antiprogestagens. Thereby Org 2058 was used as standard in both agonistic and binding assays and Org 31710 in antagonistic assays. The obtained data were compared with relative binding affinities (RBA) for both hPR-A and -B, which are present in human breast tumor MCF-7 cells, and with biopotency estimations with McPhail tests in rabbits and ovulation inhibition and pregnancy interruption tests in rats. The relative binding affinities of 14 progestagens and 7 antiprogestagens towards hPR-A, hPR-B or hPR-A/B of either CHO or MCF-7 cells were highly correlated with respect to ranking. This was also shown by the high correlation coefficients between the RBA's of hPR-B and hPR-A in CHO cells (r = 0.983) and between those of hPR-B of CHO and hPR A/B of MCF-7 cells (r = 0.957). The transactivation data of the 14 progestagens and 7 antiprogestagens for the hPR-B-MMTV-LUC cells were compared with those for hPR-A-MMTV-LUC cells and showed no differences between both cell lines with exception of the progestagens Org 32704 and 33766 and the antiprogestagen Org 33245. Therefore only the relative agonistic activities (RAA) and relative antagonistic activities (RANTA) of hPR-B-MMTV-LUC cells were compared with RBA values of hPR-B, showing a high similarity in ranking for the tested compounds, and high correlation coefficients of r = 0.91 and r = 0.97, respectively. Remarkably, RBA's were 1.6 fold higher than RAA's and RANTA's. These in vitro RBA, RAA and RANTA values for hPR-B were checked for their pharmacological relevance by in vivo biopotency measurements with the 14 progestagens and 7 antiprogestagens in rabbits and rats. The in vitro binding and transactivation potencies of progestagens appeared to be very predictive for in vivo analysis on endometrium proliferation in rabbits in the McPhail test with correlation coefficients of r = 0.81 and r = 0.87, while ovulation inhibition in rats correlated less well with r = 0.516 and r = 0.65. On the other hand, the antiprogestagenic potencies found with binding and transactivation assays had a good correlation with the potencies in the pregnancy interruption test in rats for all antiprogestagens tested, being r = 0.849 and r = 0.744, respectively. In conclusion, the binding and transactivation potencies for the tested compounds in hPR-A and -B containing cell lines showed in general a good resemblance. The transactivation studies with hPR-B-MMTV-LUC cells indicated that ranking of compounds was fairly identical to binding analysis and could be used for pre-screening of the (anti)-progestagenic bioactivity in the McPhail test in rabbits, the ovulation inhibition test and the pregnancy interruption test in rats. Therefore this transactivation assay can replace binding assays. Moreover, direct pre-screening of agonists, antagonists and partial antagonists is even possible in this in vitro bioassay, making transactivation assays for a particular class of chemical compounds to a valuable pre-screening tool for in vivo studies.
J Steroid Biochem Mol Biol 1998 Feb
PMID:Human progesterone receptor A and B isoforms in CHO cells. II. Comparison of binding, transactivation and ED50 values of several synthetic (anti)progestagens in vitro in CHO and MCF-7 cells and in vivo in rabbits and rats. 960 10

alphaA-Crystallin is a member of the small heat shock protein family that is abundantly expressed as a structural component in the vertebrate eye lens. In lenses of rodents and some other mammals, there occurs a minor variant of alphaA-crystallin, which has an insertion of 23 amino acid residues. This variant, alphaA(ins)-crystallin, results from differential integration of an optional exon into a small fraction of the mRNA. We have studied whether this alternative splicing is caused by a non-consensus cytosine in the 5' splice site adjacent to the optional exon. After replacement of the aberrant cytosine in the hamster alphaA-crystallin gene by a consensus thymine, and transient transfection of this gene in Chinese Hamster Ovary cells, the optional exon is indeed almost completely spliced into the mature mRNA. In contrast, replacement of the cytosine by adenine or guanine completely abolishes the splicing of the optional exon. Our results confirm that alternative splicing of the alphaA-crystallin primary transcript is mainly due to a non-consensus 5' splice site nucleotide. However, we conclude that the small size of the optional exon is probably an additional contributing factor and therefore it seems that the splicing mechanism is based on recognition of exons rather than introns.
Mol Biol Rep 1998 Nov
PMID:The rodent alphaA-crystallin gene: mutagenesis of a non-consensus 5'-splice site to study alternative splicing in vivo. 987 Jun 12

Androgen excess is one of the most common reproductive endocrinologic abnormalities of women. Excluding specific etiologies such as androgen-secreting neoplasms and non-classic adrenal hyperplasia, the majority of androgen excess is functional in nature. It is clear that studies concerned with the heritability of this disorder greatly depend on how it is defined. Patients with the PolyCystic Ovary Syndrome (PCOS) are clearly included. However, we argue that ovulatory women with hirsutism and hyperandrogenemia should also be considered as affected which, together with PCOS, comprise the population of women we define as having Functional Androgen Excess (FAE). Our data, and that of others, suggests that FAE/PCOS is a familial disorder, with a single autosomal dominant gene effect and a variable phenotype. Inheritance appears to be equally probable from the maternal as from the paternal side of the family. Nonetheless, our data also suggests that the affection rate among mothers is less than expected, which may be due to decreased fertility of affected mothers, or to our inability to detect the disorder in older, menopausal or hormonally treated individuals. Finally, it appears that a woman's risk for developing PCOS is approximately 40% if her sister is affected. While considering FAE/PCOS to be a dominant genetic disorder with a high degree of expressivity, its highly variable phenotype suggests that besides a single genetic mutation other factors must be contributing to the development and expression of the disorder. These factors may include environmental influences (such as fat and carbohydrate consumption) exercise level, peripubertal stress and/or hormonal exposure; and additional genetic defects, such as those that regulate insulin secretion or determine body type.
J Steroid Biochem Mol Biol
PMID:Heritability and the risk of developing androgen excess. 1041

Dopa decarboxylase converts L-dopa to dopamine, a precursor molecule for diverse biological activities in insects including neurotransmission and a variety of tanning reactions required for development, reproduction and defence against parasites. Herein, we report the cloning and sequencing of the Aedes aegypti Ddc gene, including 2.1 kb of the upstream promoter region. The transcribed region of the gene spans more than 16 kb and contains five exons. In situ hybridization localizes the blood-meal-induced ovarian transcription of this gene to the follicular epithelial cells surrounding individual oocytes. Ovary tissue transcription of Ddc is increased in response to injection of 20-hydroxyecdysone to levels equal to those observed for blood-fed controls, however coinjection with the translational inhibitor cycloheximide negates the effect, indicating an indirect regulatory role for this hormone. Clusters of putative ecdysone-responsive elements and zinc-finger binding domains for the products of Broad-Complex gene family are identified in the 5'-promoter region. These elements are discussed in the context of common insect Ddc regulatory mechanisms.
Insect Mol Biol 2000 Jun
PMID:Aedes aegypti dopa decarboxylase: gene structure and regulation. 1088 6

The aim of this study was to investigate the interaction of Stat5 with key effector proteins Erk2 and Shc after activation by growth hormone (GH), using Chinese Hamster Ovary (CHO) cells stably expressing the wild type rabbit growth hormone receptor (GHR). In coimmunoprecipitation experiments, we show GH-induced formation of complexes consisting of Stat5a and Erk2, and Stat5a and Stat5b association with the protein adaptor Shc. In CHO cells treated with GH, a rapid association of tyrosine and serine phosphorylated Stat5a with activated Erk2 is observed. In contrast, Shc proteins interact with non-phosphorylated forms of Stat5. Using truncated and tyrosine mutants of the GHR, we identify a carboxy-terminal domain of the receptor, which is critical for serine phosphorylation of Stat5a and Stat5a/Erk2 complex formation. In addition, tyrosine residues of this region of the GHR are not required for Stat5a/Erk2 interaction but are essential for Stat5a serine phosphorylation. Moreover, we detect serine phosphorylated proteins associated with Erk2, Shc and Stat5: both Stat5 isoforms interact with a serine phosphorylated protein of 63 kDa, which is shown to be related to the serine-threonine kinase Akt-1. Our results support the importance of serine phosphorylation cascades in GH signaling and open another pathway of GH signal transduction.
Mol Cell Endocrinol 2000 Aug 30
PMID:Growth hormone (GH) induces the formation of protein complexes involving Stat5, Erk2, Shc and serine phosphorylated proteins. 1099 27

This experiment was conducted to investigate the variation in lipid composition during the ovarian maturation of the crab Eriocheir sinensis. The Chinese mitten-handed crab broodstock was divided into six different maturation periods according to the size and color of ovary. Ovary, hepatopancreas, muscle, and hemolymph of broodstock in different maturation periods were analyzed for total lipid and fatty acids using gas chromatography, and lipid classes by thin-layer chromatography. The ovarian lipid concentration (expressed as percent wet ovarian weight) increased steadily from stage II (5.4%) to stage IV (19.1%), and decreased to the lowest levels after spawning (stage V, 6.6%). The hepatopancreatic lipid concentration (expressed as percent wet hepatopancreatic weight) increased with maturity of the ovaries, reached a maximum at stage III(2) (29.9%), and decreased during the subsequent period to spawning (16.7%). The muscular and hemolymph lipid concentration did not change markedly during the ovarian development. These results suggest the possible movement of hepatopancreatic lipids to the ovaries during the ovarian maturation. Both triacylglycerol and phosphatidylcholine were responsible for the increase in ovarian lipid concentration during sexual maturation. The fatty acids of total lipid, triacylglycerol, and phosphatidylcholine of the ovaries did not vary systematically during the ovarian maturation, but the ratio between n-3PUFA (polyunsaturated fatty acid) and n-6PUFA did change regularly with the ovarian lipid. These suggest that enough PUFA, especially n-3PUFA, should be supplied to the crab during ovarian maturation.
Comp Biochem Physiol B Biochem Mol Biol 2001 Aug
PMID:Variation in lipid composition of Chinese mitten-handed crab, Eriocheir sinensis during ovarian maturation. 1147 Apr 48

We investigated the effects of three components of ischemia: external acidosis (pH=6.0), extracellular hyperkalemia ([K(+)]=20 mmol/l), and resting membrane depolarization to -60 mV, on Kv4.3 current stably expressed in Chinese Hamster Ovary cells. We used single electrode whole cell patch clamp techniques to study changes in the current elicited. External acidosis caused a positive shift in the steady state activation curve from -13.4 +/- 2.1 mV to -3.3 +/- 1.5 mV (n=8, P=0.004) and the steady state inactivation curve from -56.5 +/- 0.4 mV to -46.7 +/- 0.5 mV (n=14, P<0.0001). Acidosis also caused an acceleration of recovery from inactivation with the t(1/2) decreasing from 306 ms (95% CI 287-327 ms) to 194 ms (95% CI 182-207 ms), (n=14, P<0.05). Hyperkalemia did not affect any of these parameters. Combined acidosis and hyperkalemia produced effects similar to those seen with acidosis. Changing the holding potential from -90 mV to -60 mV with test potentials of +5 and +85 mV decreased the peak currents by 34.1% and 32.4% respectively (n=14). However, in the presence of external acidosis the decrease in peak currents induced by changing the holding potential was less marked. In acidotic bath the peak current at -60 mV was reduced by only 13.6% at a test potential of +5 mV and 12.3% at a test potential of +85 mV (n=14). Taken together our data suggest that the membrane depolarization and changes in pH which occur under ischemic conditions would be accompanied by relative preservation of Kv4.3 currents and provide a molecular basis for the observation of preserved epicardial I(to) and epicardial action potential duration (APD) shortening in ischemia.
J Mol Cell Cardiol 2002 Feb
PMID:Effects of components of ischemia on the Kv4.3 current stably expressed in Chinese hamster ovary cells. 1185 59

Local immunological reactions might influence the structure of alpha1-acid glycoprotein (AGP, orosomucoid) leading to a pathological condition in e.g. the thyroid. The aim of this study was to investigate whether the AGP molecule had a direct effect on thyroid cell function in vitro. The influence of AGP and its three glycoforms, TSH (1.0 U/l), serum samples and several sugars (methyl-mannose, methyl-glycoside, N-acetyl-D-galactose, N-acetyl-D-glycoside, neuramidase) were studied with respect to their influence on the function of the Chinese Hamster Ovary (CHO) cell line transfected with the human TSH receptor (hTSHr) and on human thyroid follicular epithelial cells (TFEC) in secondary cultures. We found that low concentrations of AGP (0.001-0.05 microg/l) stimulated while high concentrations of AGP (0.25-1.0 microg/l) inhibited cAMP accumulation in both cell systems (n=24, P<0.0002). In CHO cells (JP26) and TFEC glycoforms 1 (n=9), 2 (n=12) or 3 (n=11) significantly inhibited the TSH stimulated cAMP production, respectively, compared to controls (P<0.0001) and was partially reversed by mannose (P<0.0004). Control CHO cells (JP02) without the hTSHr showed no response. The specificity of the reaction was further confirmed by binding of biotinylated glycoforms and streptavidin conjugated FITC to both cell systems. This is the first report demonstrating that AGP and/or its glycoforms affects thyroid cell function in vitro and that it does so by influencing the second messenger cAMP probably by interacting directly with the TSH receptor.
Mol Cell Endocrinol 2002 Feb 25
PMID:The influence of alpha1-acid glycoprotein (orosomucoid) and its glycoforms on the function of human thyrocytes and CHO cells transfected with the human TSH receptor. 1191 61

Partial clones for the three cynomolgus monkey (Macaca fasicularis) zona pellucida genes (cmZPA, cmZPB, and cmZPC) have previously been isolated. These partial clones contained the sequences for the C-terminal portion of each rcmZP protein. To obtain full-length clones for each cmZP, a fresh cynomolgus monkey ovarian cDNA library was constructed. PCR methodology was employed to speed the isolation of full-length clones for each cmZP cDNA. The 3' primers were designed based on sequence information from the previously identified clones; the 5' primers were designed using the human ZP sequences. The PCR technique yielded full-length clones of cmZPA and cmZPC, but not of cmZPB. Therefore, a genomic clone of cmZPB was isolated and the sequence determined. The exon/intron structure is nearly identical to the human ZPB exon/intron structure. New PCR primers were designed based on the cynomolgus monkey ZPB genomic sequence, and a full-length cmZPB cDNA was obtained. The same primers that were used to generate the cmZPB were also used to generate a baboon (Papio cynocephalus) ZPB (bZPB) cDNA. As was done previously for the human zona pellucida (hZP) cDNAs, the cmZP, and bZPB cDNAs were transferred to shuttle vectors for transfection into Chinese Hamster Ovary (CHO) cells. Stable cell lines for producing each ZP protein were isolated. Each cell line secreted the desired recombinant zona pellucida (rZP) protein into the culture medium, and each protein was purified using an established protocol. In terms of size and purity, the purified recombinant cmZP (rcmZP) and rbZPB proteins resemble the rhZP proteins.
Mol Reprod Dev 2003 Jul
PMID:Cloning and expression of cynomolgus monkey and baboon zona pellucida proteins. 1278 44

Estrogen plays an important role in the regulation of gonadotropin production in vertebrates. In this study, we isolated the estrogen receptor (ER) alpha cDNA from the goldfish pituitary. Primers for ERalpha were designed based on the similarity of selected regions (C and E domains) of known ER genes. Full-length cDNA sequence for ERalpha was determined by 3' and 5' RACE procedures. ERalpha cDNA clone was found to contain 2087 nucleotides including an open reading frame that encodes 564 amino acids, with a molecular weight of 62.8 kDa. We also cloned ERbeta-1 and ERbeta-2 from the published information and investigated the expression pattern of these ER subtypes in a variety of tissues in male and female goldfish by reverse transcriptase-polymerase chain reaction (RT-PCR). Significant variations in the relative expression of ERalpha, ERbeta-1 and ERbeta-2 were observed in different tissues in male and female goldfish. Pituitary was found to have the highest expression level of ERalpha in both male and female goldfish. Significantly, lower levels of ERalpha expression were observed in the brain, ovary, testis, liver, muscle, heart and intestine. Ovary and testis were found to have higher transcript levels of ERbeta-1 with much lower levels in the brain, pituitary, liver, muscle and heart. The ERbeta-2 was found to be expressed strongly in the pituitary followed by intestine with lower expression in other tissues. The present study provides molecular characterization of ERalpha, and information on tissue specific distribution of different ER subtypes in male and female goldfish.
Mol Cell Endocrinol 2003 Jun 30
PMID:Molecular cloning of estrogen receptor alpha and expression pattern of estrogen receptor subtypes in male and female goldfish. 1285 Feb 91


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