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1.
Summary Insulin decreased markedly the adenylyl cyclase activity associated with fat cell membranes purified by centrifugation in sucrose gradients. The hormone effect was not readily evident in crude membrane preparations.The kinetics of this effect indicate that some time was required for the onset of the insulin-induced inactivation. This lag period decreased when the insulin concentration was increased. The hormone dose dependence for adenylyl cyclase inactivation measured at a fixed time (3 min) showed a 10 to 15% decrease in activity at 1 to 30 U per ml insulin; 30 to 40% at 100 to 1000 U per ml; and 75% at 0.1 U per ml.The insulin effect was completely abolished by 0.1mm GMP-P(NH)P, 10mm fluoride, or 50 ng per ml glucagon, or by increasing the Mn++ concentration to 4mm. In addition, it was partially reversed by the addition of a fraction from the sucrose gradient, which contained soluble factors.The kinetics of the adenylyl cyclase-catalyzed reaction were studied using ATP or AMP-P(NH)P as adenylyl donor, and Mn++ or Mg++ as divalent cation, in the absence or presence of insulin. With ATP and Mg++ there was a striking reduction of the transient reaction rates after 1.5 min of incubation. Under these conditions the insulin effect was not evident. On the contrary, with ATP and Mn++ this spontaneous reduction of activity was less evident; however, in the presence of insulin there was a clear and marked reduction of the transient reaction rate measured after 1.5 min of incubation. With AMP-P(NH)P the kinetic data were qualitatively similar to those observed with ATP.It is concluded that under certain assay conditions adenylyl cyclase may be converted to an inactive enzyme form, and that such a conversion is more evident in the presence of Mg++ than with Mn++. In the latter case, insulin appears to enhance the rate of this conversion. 相似文献
2.
W. Plenert 《Histochemistry and cell biology》1959,1(3):171-177
Zusammenfassung Die Technik des Nachweises der a. L.-Ph. in Blutausstrichen wird besprochen. Zur Fixierung wird die Methode vonKaplow empfohlen: 90% Methanol, 10% Formol 1:10, 30 sec bei 0°C. Vergleichswerte mit anderen Fixantien werden aufgeführt.Metallionen aktivieren die a. L.-Ph. in derart fixierten Ausstrichen in der Reihenfolge abnehmender Wirksamkeit:Mg++-Fe+++-Co++-Mn++-Cu++-Fe++. Die Wirkung aller Ionen erwies sich als stark konzentrationsabhängig. Ni++, Zn++ und Pb++ hemmten ebenfalls konzentrationsabhängig.In den Blutausstrichen ist die Spaltungsgeschwindigkeit des sauren Na--Naphthylphosphats (Azo-Kupplung) weit höher als die des -Glycerophosphats bei maximaler Mg++-Aktivierung mit der Calcium-Kobalt-Methode nachGomori-Takamatsu. Die Spaltungsgeschwindigkeit von -Glycerophosphat kann durch Zusetzen kleiner Mengen Fe+++ und Cu++ über die Mg++-Aktivierung hinaus gesteigert werden bei Beschleunigung der Anfangsgeschwindigkeit der Hydrolyse.
Mit 2 Textabbildungen 相似文献
Summary A technic is described for demonstrating the activity of alkaline phosphatase in human leucocytes. For fixation the method ofKaplow is recommended: 90 per cent methanol +10 per cent formalin 1:10 for 30 seconds at 0°C. Other fixatives are evaluated.Under the conditions of the experiment metal ions activated the alkaline phosphatase of leucocytes in the following order: Mg++>Fe+++>Co++>Mn++>Cu++>Fe++.The effectiveness of all these metal ions was to a high degree dependent on concentration. Ni++, Zn++, and Pb++ showed an inhibitory effect also dependent on concentration.In fixed blood smears the velocity of hydrolysis of sodium--naphthylphosphate (simultaneous azo-coupling technic) is far greater than that of Na--glycerophosphate (Ca-Co-method of Gomori-Takamatsu). Adding small amounts of Fe+++ and Cu++ to the incubation medium, it is possible to increase the velocity of hydrolysis of glycerophosphate beyond the values of maximal Mg++ activation and simultaneously enhancing the initial velocity of the reaction.
Mit 2 Textabbildungen 相似文献
3.
Summary When grown with glucose, S. discophorus synthesized large amounts of poly--hydroxybutyrate which accumulated intracellularly as sudanophilic granules. The rate of endogenous oxygen consumption by such cells was markedly increased by Mn++ and even more by Mg++. It has been shown that these inorganic ions stimulate the oxidation of the intracellular poly--hydroxybutyrate.Dedicated by the senior author to Prof. C. B. van Niel on the occasion of his 70th birthday with gratitude for many unforgettable years of association, instruction and stimulation. 相似文献
4.
Properties of -glucan synthetase from S. cerevisiae were studied. The enzyme exhibited optimal activity at pH 6.7 and 24 C. Km for UDP-glucose was 0.12mm. Addition of Mg++ or Mn++ stimulated its activity by 60% and 21% respectively. High concentrations of EDTA and hydroxyquinoline were inhibitory. Glucan synthetase was fully active in cell-free extracts. Small concentrations of trypsin or subtilopeptidase A from Bacillus subtilis, caused only a slight increase in glucosyl transferase activity, but larger concentrations destroyed -glucan synthetase. Acid proteases were neither stimulatory nordestructive. Thus it seemsunlikelythat -glucan synthetase exists in a zymogen form. Glucan synthetase was unstable. It was inactivated more rapidly at 28 C than at 0 C. The presence of substrate, -glucan or the protease inhibitors PMSF, Antipain or Pepstatin A did not protect -glucan synthetase from inactivation. Glucan synthetase was not stimulated by addition of cellobiose or -glucans. The synthesis of -glucans was competitively inhibited by UDP (Ki=0.45mm). Glucono--lactone, a known inhibitor of -glucosidases was a strong non-competitive inhibitor of -glucan synthetase.This work was supported by grants PNCB 00071 and 847 of the Consejo Nacional de Ciencia y Tecnología, México. 相似文献
5.
A number of N- and C-terminal deletion and point mutants of bovine -1,4 galactosyltransferase (-1,4GT) were expressed inE. coli to determine the binding regions of the enzyme that interact withN-acetylglucosamine (NAG) and UDP-galactose. The N-terminal truncated forms of the enzyme between residues 1–129, do not show any significant difference in the apparentK
ms toward NAG or linear oligosaccharide acceptors e.g. for chitobiose and chitotriose, or for the nucleotide donor UDP-galactose. Deletion or mutation of Cys 134 results in the loss of enzymatic activity, but does not affect the binding properties of the protein either to NAG- or UDP-agarose. From these columns the protein can be eluted with 15mm NAG and 50mm EDTA, like the enzymatically active protein, TL-GT129, that contains residues 130–402 of bovine -1,4GT. Also the N-terminus fragment, TL-GT129NAG, that contains residues 130–257 of the -1,4GT, binds to, and elutes with 15mm NAG and 50mm EDTA from the NAG-agarose column as efficiently as the enzymatically active TL-GT129. Unlike TL-GT129, the TL-GT129NAG binds to UDP-columns less efficiently and can be eluted from the column with only 15mm NAG. The C-terminus fragment GT-257UDP, containing residues 258–402 of -1,4GT, binds tightly to both NAG- and UDP-agarose columns. A small fraction, 5–10% of the bound protein, can be eluted from the UDP-agarose column with 50mm EDTA alone. The results show that the binding behaviour of N- and C-terminal fragments of -1,4GT towards the NAG- and UDP-agarose columns differ, the former binds preferentially to NAG-columns, while the latter binds to UDP-agarose columns via Mn2+. 相似文献
6.
Summary To study Cl conductive and cotransport mechanisms, primary cultures of canine tracheal cells were grown to confluency on thin glass cover slips and on porous filters. Transepithelial resistance was >100 ·cm2, and short circuit current (I
sc=2–20 A/cm2), representing active secretion of Cl, increased >threefold with addition of 10 m isoproterenol to the serosal solution. Cells made transiently permeable in hypotonic solution were loaded with the Cl-sensitive fluorophore 6-methoxy-N-(3-sulfopropyl) quinolinium (SPQ) (5mm, 4 min, 150 mOsm). The electrical properties of the cell monolayers were not altered by the loading procedure. Intracellular SPQ fluorescence was monitored continuously by epifluorescence microscopy (excitation 360±5 nm, emission>410 nm). SPQ leakage from the cells was <10% in 60 min at 37°C. Intracellular calibration of SPQ fluorescencevs. [Cl] (0–90mm) was carried out using high-K buffers containing the ionophores nigericin (5 m) and tributyltin (10 m); SPQ fluorescence was quenched with a Stern-Volmer constant of 13m
–1. Intracellular Cl activity was 43±4mm. Cl flux was measured in response to addition and removal of 114mm Cl from the bathing solution. Addition of 10 m isoproterenol increased Cl efflux from 0.10 to 0.27mm/sec. The increase was inhibited by the Cl-channel blocker diphenylamine-2-carboxylic acid (1mm). In the absence of isoproterenol, removal of external Na or addition of 0.5mm furosemide, reduced Cl influx by >fourfold. In ouabain-treated monolayers, removal of external K in the presence of 5mm barium diminished Cl influx by >twofold, suggesting that Cl entry is in part K dependent. These results establish an accurate optical method for the realtime measurement of intracellular Cl activity in tracheal cells that does not require an electrically tight cell monolayer. The data demonstrate the presence of an isoproterenol-regulated Cl channel and a furosemide-sensitive cation-coupled transport mechanism. 相似文献
7.
Summary Calcium markedly stimulates the respiration of the isolated chick chorioallantoic membrane. This stimulation of oxygen uptake appears to be closely associated with the membrane's active transcellular calcium transport mechanism. In the presence of 1mm Ca++ the rate of uptake increases from 9.3±0.15 to 13.0±0.2 liters O2/cm2/hr, an increase of about 40%. The calcium-stimulated respiration is specific for the ectodermal layer of cells, the known location of the calcium transport mechanism, and only occurs when the calcium transport mechanism is operative. Sr++ and Mn++ are transported by the tissue at a lower rate than Ca++ and cause a smaller stimulation of oxygen consumption. Mg++ and La3+ have no effect on tissue respiration. In the presence of Ca++, the organic mercurialp-chloromercuribenzene sulfonate (PCMBS) inhibits calcium transport and specifically decreases the oxygen uptake of the ectoderm to a rate identical to that obtained in a calcium-free medium. Stripping the inner shell membrane away from the chorioallantoic membrane mimics these effects. The specificity and locus of action of these two inhibitors suggest that a vital component of the active transcellular calcium transport mechanism resides on or near the outer surface of the plasma membrane of the ectodermal cells and that sulfhydryl groups are important to the normal function of this component. 相似文献
8.
M. J. Avison S. R. Gullans T. Ogino G. Giebisch 《The Journal of membrane biology》1988,105(3):197-205
Summary Addition of glucose or the nonmetabolizable analogue -methyl-d-glucoside to rabbit proximal tubules suspended in a glucoseand alanine-free buffer caused a sustained increase in intracellular Na+ content (+43±7 nmol · (mg protein)–1) and a concomitant but larger decrease in K+ content (–72±11 nmol· (mg protein)–1). A component of the net K+ efflux was Ba2+ insensitive, and was inhibited by high (1mm) but not low (10 m) concentrations of the diuretics, furosemide and bumetanide. The increase in intracellular Na+ content is consistent with the view that the increased rates of Na+ and water transport seen in the proximal tubule in the presence of glucose can be attributed (at least in part) to a stimulation of basolateral pump activity by an increased [Na+]
i
. 相似文献
9.
Summary The Ca++-mediated increase in K+-permeability of intact red blood cells (Gardos effect) was initiated by exposing cells to known concentrations of Ca++ (using EGTA buffers) in the presence of the ionophore A23187. The potency of quinine, an inhibitor of the response, was found to depend on the external K+ concentration. In K+-free solutions the concentration of quinine to achieve 50% inhibition (K
50) was 5 m, but at 5mm K+ the required concentration was increased 20-fold to 100 m. An increase in internal Na+ had the opposite effect, allowing a high potency of quinine despite the presence of external K+. Alterations in the internal K+ level, on the other hand, were without effect on theK
50, suggesting that the membrane potential is not a factor. This conclusion is supported by the lack of effect on quinine inhibition of substitution of Cl– by NO
3
–
, a considerably more permeant anion. The data are consistent with the hypothesis that quinine inhibits by competitively displacing K+ from an external binding site, the reported K+-activation site for the Ca++-mediated K+-permeability. 相似文献
10.
Summary The characteristics of the cholera toxin-stimulated adenylate cyclase of toad (Bufus marinus) and rat erythrocyte plasma membranes have been examined, with special emphasis on the response to purine nucleotides, fluoride, magnesium and catecholamine hormones. Toad erythrocytes briefly exposed to low concentrations of cholera toxin (40,000 to 60,000 molecules per cell) and incubated 2 to 4 hr at 30°C exhibit dramatic alterations in the kinetic and regulatory properties of adenylate cyclase. The approximateK
m
for ATP, Mg++ increases from about 1.8 to 3.4mm in the toxinstimulated enzyme. The stimulation by cholera toxin increases with increasing ATP, Mg++ concentrations, from 20% at low levels (0.2mm) to 500% at high concentrations (greater than 3mm). Addition of GTP, Mg++ (0.2mm) restores normal kinetic properties to the toxin-modified enzyme, such that stimulation is most simply explained by an elevation ofV
max. GTP enhances the toxin-treated enzyme activity two-to fourfold at low ATP concentrations, but this effect disappears at high levels of the substrate. At 0.6mm ATP and 5mm MgCl2 the apparentK
a
for GTP, Mg++ is 5 to 10m. The control (unstimulated) enzyme demonstrates a very small response to the guanyl nucleotide. 5-ITP also stimulates the toxin-treated enzyme but cGMP, guanine, and the pyrimidine nucleotides have no effect. Cholera toxin also alters the activation of adenylate cyclase by free Mg++, decreasing the apparentK
a
from about 25 to 5mm. (–)-Epinephrine sensitizes the toad erythrocyte adenylate cyclase to GTP and also decreases the apparentK
a
for free metal. Sodium fluoride, which cause a 70- to 100-fold activation of enzyme activity, has little effect on sensitivity to GTP, and does not change the apparentK
a
for Mg++; moreover, it prevents modulation of these parameters by cholera toxin. Conversely, cholera toxin severely inhibits NaF activation, and in the presence of fluoride ion the usual three- to fivefold stimulation by toxin becomes a 30 to 60% inhibition of activity. The toxin-stimulated enzyme can be further activated by catecholamines; in the presence of GTP the (–)-epinephrine stimulation is enhanced by two- to threefold. The increased catecholamine stimulation of toad erythrocyte adenylate cyclase induced by cholera toxin is explained primarily by an increase in the maximal extent of activation by the hormones. Rat erythrocyte adenylate cyclase is also modified by cholera toxin. In the mammalian system the apparent affinity for the hormone appears to be increased. Cholera toxin thus induces profound and nearly permanent changes in adenylate cyclase by a unique process which mimics the stimulation by hormones in important ways, and which also accentuates the normal hormonal response. The relevance of these findings to the mechanism of action of cholera toxin is considered.Part of this work was reported at the 1974 meeting of the Federation of American Societies for Experimental Biology (Bennett & Cuatrecasas, 1974). 相似文献
11.
The effect of divalent metal ions on the activity of Mg++ depleted ribulose-1,5-bisphosphate oxygenase 总被引:1,自引:0,他引:1
Ribulose-1,5-bisphosphate carboxylase-oxygenase is deactivated by removal of Mg++. The enzyme activities can be restored to a different extent by the addition of various divalent ions in the presence of CO2. Incubation with Mg++ and CO2 restores both enzyme activities, whereas, the treatment of the enzyme with the transition metal ions (Mn++, Co++, and Ni++) and CO2 fully reactivates the oxygenase: however, the carboxylase activity remains low. In experiments where CO2-free conditions were conscientiously maintained, no reactivation of RuBP oxygenase was observed, although Mn++ ions were present. Other divalent cations such as Ca++ and Zn++, restore neither the carboxylase nor the oxygenase reaction. Furthermore, the addition of Mn++ to the Mg++ and CO2 preactivated enzyme significantly inhibited carboxylase reactions, but increased the oxygenase reaction.Abbreviation RuBP
ribulose-1,5-bisphosphate. The enyme unit for RuBP carboxylase is defined as mol CO2 fixed·min-1 and for the RuBP oxygenase as mol O2 consumed · min-1 相似文献
12.
Glycerol kinase is induced in cells ofPseudomonas aeruginosa strain PAO when grown in the presence of glycerol or glycerol-3-phosphate. The enzyme was isolated from the soluble cytoplasmic fraction of cell extracts and purified 500-fold by ammonium sulfate precipitation and chromatography on columns of Sephadex G-25, DEAE-Sephadex, hydroxyapatite, and Sephadex G-200. A molecular weight of 120,000 was estimated by gel filtration of the catalytically active enzyme. In polyacrylamide gel electrophoresis the purified product contained one major band of Coomassie Blue staining material. The enzyme exhibited an apparent Km of 40 M for glycerol and 23 M for ATP. Of the nucleotide triphosphates tested, only ATP served as a phosphoryl group donor. Mg++ or Mn++ was required for activity, although a threefold greater concentration of Mn++ was required when Mn++ substituted for Mg++. In contrast to most other catabolic glycerol kinases in bacteria, the enzyme was not inhibited by fructose-1,6-diphosphate nor by other tested metabolites. 相似文献
13.
The first enzyme (named GTP cyclohydrolase) in the pathway for the biosynthesis of pteridines has been partially purified from extracts of late pupae and young adults of Drosophila melanogaster. This enzyme catalyzes the hydrolytic removal from GTP of carbon 8 as formate and the synthesis of 2-amino-4-hydroxy-6-(d-erythro-1,2,3-trihydroxypropyl)-7,8-dihydropteridine triphosphate (dihydroneopterin triphosphate). Some of the properties of the enzyme are as follows: it functions optimally at pH 7.8 and at 42 C; activity is unaffected by KCl and NaCl, but divalent cations (Mg2+, Mn2+, Zn2+, and Ca2+) are inhibitory; the K
m for GTP is 22 m; and the molecular weight is estimated at 345,000 from gel filtration experiments. Of a number of nucleotides tested, only GDP and dGTP were used to any extent as substrate in place of GTP, and these respective compounds were used only 1.8% and 1.5% as well as GTP.This work was supported by research grants from the National Institutes of Health (AM03442) and the National Science Foundation (GB33929). 相似文献
14.
Summary Both simultaneous and consecutive mechanisms for Na+–Ca++ exchange are formulated and the associated systems of steady-state equations are solved numerically, and the net and unidirectional Ca++ fluxes computed for a variety of ionic and electrical boundary conditions. A simultaneous mechanism is shown to be consistent with a broad range of experimental data from the squid giant axon, cardiac muscle and isolated sarcolemmal vesicles. In this mechanism, random binding of three Na+ ions and one Ca++ on apposing sides of a membrane are required before a conformational change can occur, translocating the binding sites to the opposite sides of the membranes. A similar (return) translocation step is also permitted if all the sites are empty. None of the other states of binding can undergo such translocating conformational changes. The resulting reaction scheme has 22 reaction steps involving 16 ion-binding intermediates. The voltage dependence of the equilibrium constant for the overall reaction, required by the 31 Na+Ca++ stoichiometry was obtained by multiplying and dividing, respectively, the forward and reverse rate constants of one of the translocational steps by exp(–FV/2RT). With reasonable values for the membrane density of the enzyme (120 sites m2) and an upper limit for the rate constants of both translocational steps of 105·sec–1, satisfactory behavior was obtainable with identical binding constants for Ca++ on the two sides of the membrane (106
m
–1), similar symmetry also being assumed for the Na+ binding constant (12 to 60m
–1). Introduction of order into the ion-binding process eliminates behavior that is consistent with experimental findings. 相似文献
15.
Summary NADH oxidation with the particulate fraction from dark aerobically grown Rhodospirillum rubrum is significantly stimulated by the addition of phosphate (Pi) and Mg++, or Pi, Mg++, ATP and the hexokinase-glucose system. K
m
values for Pi in NADH oxidation and phosphorylation are 10–3
m and 8×10–4
m, respectively. These K
m
values are almost the same as in corresponding photophosphorylation and oxidative phosphorylation catalyzed with chromatophores. As in the case of NADH oxidation with chromatophores, NADH oxidation with the particulate fraction has an optimal pH at 7.5 without additions, which is shifted to 6.9 by the addition of Pi and Mg++, or Pi, Mg++, ATP and the hexokinase-glucose system. The optimal pH for coupled phosphorylation is 6.9. 10 g per ml of oligomycin can suppress stimulation of NADH oxidation by Pi, or by the energy trapping system, and prevent the shift of optimal pH. The particulate fraction can catalyze Pi-incorporation into glucose-6-phosphate without externally added ATP, so that Pi-incorporation is inhibited by oligomycin. From these findings, it is concluded that NADH oxidation in the particulate fraction is tightly coupled to phosphorylation. 相似文献
16.
In cell-free patches from cultured neonatal rat cardiocytes, the cytosolic presence of GTP--S (100 µmol/l) or GDP-\-S (100 µmol/1) activated K(ATP)
+ channels. GTP--S required cytosolic Mg++, suggesting that an activated G-protein causes the increase in open probability. The great variations of the channel response to GTP--S and GDP-\-S indicates that cardiac K(ATP)
+ channels represent a heterogeneous family.
Correspondence to: M. Kohlhardt 相似文献
17.
Jeffrey L. Garvin Sidney A. Simon Edward J. Cragoe Jr. Lazaro J. Mandel 《The Journal of membrane biology》1985,87(1):45-54
Summary Several new amiloride analogues and two reported photoaffinity analogues were tested for irreversible inhibition of short-circuit current,I
sc, in toad bladder. Bromoamiloride, a photoaffinity analogue, induced 40% irreversible inhibition at 500 m after irradiation with ultraviolet light 320 nm. Iodoamiloride caused no irreversible inhibition. Of the new analogues tested, only 3,5-diamino-6-chloro-N-[(phenylamino) aminomethylene] pyrazinecarboxamide,phenamil, irreversibly inhibitedI
sc at concentrations of 0.05 to 5 m when added to the mucosal solution. Irreversible inhibition ofI
sc by phenamil may be attributed to specific blockage of the mucosal sodium channels, which depended on: 1) time of exposure; 2) mucosal pH: 3) mucosal sodium concentration. For example, 5 m phenamil irreversibly inhibitedI
sc by 38% in 103mm Na at pH 8.6 and nearly 75% in 30mm Na at pH 6.4 after a 40-min exposure. Irreversible inhibition occurred in two phases with time constants of 10 min and approximately 140 min. Due to its irreversible nature, phenamil may be used to measure channel density. 相似文献
18.
Santiago LLovera Ramon Bonet Maria Dolores Simon-Pujol Francisco Congregado 《Applied microbiology and biotechnology》1993,39(3):424-426
The influence of some ions in pre-growth culture medium on chromate reduction by resting cells of Agrobacterium radiobacter strain EPS-916 was investigated. The reduction was dependent on the Fe2+ content of the culture medium: the higher the iron content, the lower the reduction rate. The cells showed maximum chromate reduction when pre-grown in the presence of 0.243 m Mg2+, 20 m Ca2+ and 3.6 m Mn2+. Chromate reduction was not affected by the addition of MgCl2, CdCl2, ZnCl2, MnCl2, Na2SO4 (1000 m), and Na2MoO4 (100 m) to the activity assays. However, activity was inhibited by the presence of Na2SO4 (10 mm), Na2MoO4 (200 m) and ferric citrate. 相似文献
19.
M. M. Sorenson J. P. Reuben A. B. Eastwood M. Orentlicher G. M. Katz 《The Journal of membrane biology》1980,53(1):1-17
Summary Precipitation of Ca oxalate in the sarcoplasmic reticulum of chemically skinned rabbit psoas fibers caused an increase in light scattering which was proportional to the amount of Ca accumulated per unit fiber volume. The increase in scattering was used to measure net accumulation rates and steady-state Ca capacities of the sarcoplasmic reticulum in single fibers. The data obtained were qualitatively and quantitatively similar to those reported for isolated vesicle preparations.Under conditions in which Ca was not depleted from the medium, Ca accumulation was linear with time over much of its course. Steady-state capacities were independent of the Ca concentration; uptake rates were half-maximal at 0.5 m Ca++ and saturated above about 1.0 m. Both rate and capacity varied with the oxalate concentration, being maximal at oxalate concentrations >=5mm and decreasing in proportion to one another at lower concentrations, with a threshold near 0.25mm. At the lower loads, electron micrographs showed many sarcoplasmic reticulum elements empty of precipitate alongside others that were full, whereas virtually all were filled in maximally loaded fibers. These data indicate that the Ca oxalate capacity of each fiber varies with the number and volume of elements in which Ca oxalate crystals can form at a given oxalate concentration, and that individual regions of the sarcoplasmic reticulum within each sarcomere differ in their ability to support Ca oxalate precipitation. Our working hypothesis is that this range in ability to form Ca oxalate crystals involves differences in ability to accumulate and retain ionized Ca inside the sarcoplasmic reticulum. 相似文献
20.
In cyanobacteria, the glutamine synthetase-L-glutamine-2-oxoglutarate aminotransferase (GS-GOGAT) pathway is the major ammonia-assimilating route. The GS ofAnabaena doliolum was synthesized more under N2-fixing conditions, followed by ammonium, nitrate, and nitrite as nitrogen sources. The activities of both the glutamine synthetase, Mg2+-dependent biosynthetic and Mn2+-dependent -glutamyl transferase were optimum at pH 7. The active site of the enzyme bears sulfhydryl (-SH) groups; this was confirmed with the-SH group inhibitors, para-chloromercuribenzoate (pCMB) and N-ethylmaleimide (NEM). The biosynthetic and -glutamyl transferase activities showed specificity for the divalent cations, Mg2+ and Mn2+, respectively. The other divalent cations Co2+, Cu2+, and Ni2+ were poor substitutes. This enzyme also required these divalent cations to stabilize its structure and function under extreme conditions such as high and low temperatures and urea denaturation. The glutamate analogl-methionine-d,l-sulfoximine, inactivated the enzyme, whereas the GOGAT inhibitor, azaserine, had no effect on the enzyme activity. The GS enzyme required de novo protein synthesis. 相似文献