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51.
Does the guanine nucleotide regulatory protein Ni mediate progesterone inhibition of Xenopus oocyte adenylate cyclase? 下载免费PDF全文
In Xenopus laevis oocytes progesterone is able to inhibit directly the plasma membrane adenylate cyclase activity and induce reinitiation of meiotic maturation. To determine whether progesterone inhibition is mediated by the inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase, Ni, the effect of the Bordetella pertussis toxin (IAP) and limited proteolysis on progesterone action in oocytes was investigated. Treatment of oocyte membranes with islet activating protein (IAP) in the presence of [32P]NAD led to incorporation of radiolabel into a 41 000-dalton membrane protein. However, exposure of isolated oocytes to 100 ng/ml IAP for up to 24 h, or oocyte membranes with concentrations of toxin as high as 100 micrograms/ml, had no effect on either progesterone inhibition of adenylate cyclase or induction of maturation. Similarly, limited alpha-chymotrypsin proteolysis of oocyte membranes failed to modify progesterone-induced inhibition of adenylate cyclase. In contrast, inhibition of human platelet adenylate cyclase by epinephrine, acting via a GTP-dependent, alpha 2-adrenergic receptor-mediated pathway, is almost completely abolished by both IAP treatment and limited proteolysis of platelet membranes. These data indicate that unlike attenuation of platelet enzyme activity, the inhibition of adenylate cyclase in oocyte membranes by progesterone does not occur via a classical Ni-mediated pathway. 相似文献
52.
Antibodies against highly purified B-subunit of the chick oviduct progesterone receptor 总被引:5,自引:0,他引:5
P Tuohimaa J M Renoir C Radanyi J Mester I Joab T Buchou E E Baulieu 《Biochemical and biophysical research communications》1984,119(2):433-439
A rabbit was immunized with the highly purified B-subunit (110kDa) (20 to 50 micrograms per injection) of the chick oviduct progesterone receptor (PR). Specific antibodies (IgG-RB) were observed 2 weeks after the first booster injection and high antibody titers in the serum were found after the second and third booster injections (with Kdeq of interaction integral of 2 nM). IgG-RB were tested by immunoprecipitation, immunoblotting, density gradient ultracentrifugation and protein A-sepharose assay methods. They recognized not only the B-subunit but also the A-subunit (79K), the nuclear PR, the mero-receptor (proteolytic cleavage product) and the "non-activated" molybdate-stabilized "8S" PR. However, IgG-RB did not interact with the 90K non hormone-binding component of this 8S-PR. IgG-RB did not affect the binding of the hormone to PR, whether incubated with the receptor before or after labelling with tritiated progesterone. They did not cross-react with glucocorticosteroid receptor of the chick oviduct. Weak interaction was observed with estrogen receptor of the chick oviduct and with KC1 activated "4S" forms of the rabbit and human uterus PR. 相似文献
53.
R D Nadler C Roth-Meyer J Wallis W Cooper E Bourreau G Affre E E Baulieu 《Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie》1984,298(14):409-413
Ten male and eleven female Chimpanzees from three to nine years of age were studied to establish possible correlations between behavioral changes and hormonal changes peculiar to adrenarche and the pre-puberty period. Most of the thirteen behaviors studied varied with age, body weight and hormones. For the males, the correlations were significant statistically for age, weight and plasma concentration of testosterone and of FSH. The correlations for the females were more often not significant statistically. In ten out of the thirteen behaviors for the female, however, the correlations with the sulfate of plasma dehydroepiandrosterone were in the same direction as those observed for the males with testosterone. 相似文献
54.
William B. Guggino Emile L. Boulpaep Gerhard Giebisch 《The Journal of membrane biology》1982,65(3):185-196
Summary The chloride conductance of the basolateral cell membrane of theNecturus proximal tubule was studied using conventional and chloride-sensitive liquid ion exchange microelectrodes. Individual apical and basolateral cell membrane and shunt resistances, transepithelial and basolateral, cell membrane potential differences, and electromotive forces were determined in control and after reductions in extracellular Cl–. When extracellular Cl– activity is reduced in both apical and basolateral solutions the resistance of the shunt increases about 2.8 times over control without any significant change in cell membrane resistances. This suggests a high Cl– conductance of the paracellular shunt but a low Cl– conductance of the cell membranes. Reduction of Cl– in both bathing solutions or only on the basolateral side hyperpolarizes both the basolateral cell membrane potential difference and electromotive force. Hyperpolarization of the basolateral cell membrane potential difference after low Cl– perfusion was abolished by exposure to HCO
3
–
-free solutions and SITS treatment. In control conditions, intracellular Cl– activity was significantly higher than predicted from the equilibrium distribution across both the apical and basolateral cell membranes. Reducing Cl– in only the basolateral solution caused a decrease in intracellular Cl–. From an estimate of the net Cl– flux across the basolateral cell membrane and the electrochemical driving force, a Cl– conductance of the basolateral cell membrane was predicted and compared to measured values. It was concluded that the Cl– conductance of the basolateral cell membrane was not large enough to account for the measured flux of Cl– by electrodiffusion alone. Therefore these results suggest the presence of an electroneutral mechanism for Cl– transport across the basolateral cell membrane of theNecturus proximal tubule cell. 相似文献
55.
Influence of the position of the double bond in steroid substrates on the efficiency of the proton-transfer reaction by Pseudomonas testosteroni 3-oxo steroid Δ4–Δ5-isomerase 下载免费PDF全文
Studies of the proton-transfer reaction by Pseudomonas testosteroni 3-oxo steroid Delta(4)-Delta(5)-isomerase with Delta(5(6))- and Delta(5(10))-steroid substrates demonstrate the importance of the position of the double bond for the efficiency of the isomerization process. Thus 3-oxo-Delta(5(6))-substrates have markedly high k(cat.) values, whereas those of 3-oxo-Delta(5(10))-substrates are very low and their apparent K(m) values approach equilibrium dissociation constants. The first step in the isomerization process is: [Formula: see text] which is governed by the k(-1)/k(+1) ratio and is shown to be very similar for the two classes of substrates (3-oxo-Delta(5(6))- and -Delta(5(10))-steroids). They therefore differ in the steps distal to the initial formation of the Michaelis-Menten complex. The use of the deuterated androst-5(6)-ene-3,17-dione substrate enabled us to calculate individual rate constants k(+1) and k(-1) as well as to determine the apparent rate-limiting step in the isomerization process. With the deuterated oestr-5(10)-ene-3,17-dione substrate, no significant isotope effect was observed suggesting that a different rate-limiting step may be operative in this isomerization process. Data are presented that indicate that under optimal concentrations of the efficient androst-5(6)-ene-3,17-dione substrate, the forward reaction for ES complex formation (as defined by k(+1)) is limited only by diffusion and the apparent K(m) does not approach the equilibrium constant, suggesting that the evolution of this enzyme has proceeded close to ;catalytic perfection'. 相似文献
56.
E.V. Younglai F. Godeau J. Mester E.E. Baulieu 《Biochemical and biophysical research communications》1980,96(3):1274-1281
The activity of ornithine decarboxylase (ornithine carboxylyase E.C. 4.1.1.17) was studied during meiotic maturation induced in vitro by progesterone in follicle cell-free oocytes. Enzyme activity increased 4–6 fold during maturation, preceding germinal vesicle breakdown. The increase in ornithine decarboxylase activity was inhibited by cholera toxin, an agent that blocks meiotic maturation and increases cAMP levels within the cell. It was also prevented by cycloheximide but not by actinomycin D. Treatment of oocytes with D,L-α-difluoromethyl-ornithine, an irreversible inhibitor of ornithine decarboxylase and of putrescine synthesis, effectively abolished enzyme activity without preventing germinal vesicle breakdown. These observations show that the progesterone-induced increase in ornithine decarboxylase activity is not required for completion of meiotic division of the oocyte. 相似文献
57.
Willy J. Malaisse Kowluru Anjaneyulu Renu Anjaneyulu Etienne Couturier 《Molecular and cellular biochemistry》1980,30(2):67-70
Summary In an artificial system, the ionophore A23187, which transports Ca2+ but not Na+, is able to mediate the uphill translocation of Ca2+ from one aqueous medium to another across an organic immiscible phase, provided that a Na+, Li+ or H+ gradient is imposed on the system. Therefore, in the process known as Na-Ca countertransport, the downhill influx of Na+ may not be necessary for causing Ca2+ extrusion against its electrochemical gradient. 相似文献
58.
The antibiotic ionophore ionomycin translocates Ca from an aqueous medium into or across an organic immiscible phase. At pH 8.0, ionomycin translocates less Ca than A23187, the effects of these ionophores being additive to one another. The capacity of ionomycin to translocate Ca across the organic phase is dramatically decreased when the pH of the aqueous media is reduced from 8.0 to 7.5 or lower values. Ionomycin also mediates Ca exchange-diffusion in liposomes, the magnitude of such a process being greater in fluid than in rigid liposomes. At a physiological pH (7.4), ionomycin is unexpectedly as potent as A23187 in mediating Ca transport in fluid liposomes. These findings suggest that the capacity of ionophores to translocate Ca across model membranes depends on both the transverse and lateral mobility of the ionophoretic molecules. The relative importance of the latter phenomenon itself largely depends on the stoichiometry of the Ca-ionophore complex. 相似文献
59.
60.
Josette Martin-Tanguy Ferdinand Cabanne Etienne Perdrizet Claude Martin 《Phytochemistry》1978,17(11):1927-1928
Hydroxycinnamic acid amides have been identified as the main phenolic constituents in the reproductive organs of a range of flowering plants. 相似文献