共查询到20条相似文献,搜索用时 15 毫秒
1.
Iu I Gubski? R G Primak L A Bodanova A G Goriushko O V Zadorina M I Golubov 《Ukrainski? biokhimicheski? zhurnal》1991,63(4):87-92
Physico-chemical parameters of membranes of skeletal muscles' sarcoplasmic reticulum in antioxidant insufficiency, which was modelled by excluding alpha-tocopherol from the animals ration, and after treatment with phenol antioxidant ionol were studied. It was shown that activation of lipid peroxidation in vitamin E insufficiency results in a significant lowering of microviscosity of lipid bilayer membranes of sarcoplasmic reticulum. Using polarography significant changes in membrane protein conformation were revealed, which were characterized by lowering of integrity and by disorganization of protein globules. Treatment of animals with antioxidant insufficiency with ionol led to certain normalization of changes of physico-chemical characteristics of the learned membrane structures caused by lipid peroxidation. 相似文献
2.
Calcium releasing action of quercetin on sarcoplasmic reticulum from frog skeletal muscle 总被引:1,自引:0,他引:1
The release of Ca by quercetin from the sarcoplasmic reticulum has been claimed to be a result of the well-known inhibition of Ca2+-ATPase activity, or to be due to an intrinsic property of quercetin. To get a clearer understanding of the effect of quercetin, we examined it using fragmented sarcoplasmic reticulum (FSR) from bullfrog skeletal muscle. The rapid phase of Ca release (hereafter simply referred to as "Ca release") from loaded FSR was almost completed within 5 s after addition of quercetin in the presence of ATP. It cannot be ascribed to the inhibition of Ca2+-ATPase activity on the basis of following findings. First, when Ca uptake was driven by carbamylphosphate, no or little Ca release was observed in marked contrast to a stronger reduction in the rate of Ca uptake. Secondly, procaine reverses the Ca releasing action of quercetin, whereas it show a synergistic action in the inhibition of Ca2+-ATPase activity. Thirdly, HFSR released more Ca than LFSR, while the Ca2+-ATPase activities of both fractions were inhibited to a similar extent. The Ca release by quercetin is enhanced by ATP or beta, gamma-methylene adenosine triphosphate, and decreased by procaine or a high concentration of Mg2+. In the presence of 2.5 mM caffeine, the amount of Ca2+ released by quercetin was decreased, and the dose-effect relationship was shifted to higher doses of quercetin. This indicates that quercetin and caffeine probably overlap in the site(s) of the action, but that quercetin is dissimilar from halothane in the mode of its Ca-releasing action. 相似文献
3.
To define the mechanism responsible for the slow rate of calcium transport by cardiac sarcoplasmic reticulum, the kinetic properties of the Ca2+-dependent ATPase of canine cardiac microsomes were characterized and compared with those of a comparable preparation from rabbit fast skeletal muscle. A phosphoprotein intermediate (E approximately P), which has the stability characteristics of an acyl phosphate, is formed during ATP hydrolysis by cardiac microsomes. Ca2+ is required for the E approximately P formation, and Mg2+ accelerates its decomposition. The Ca2+ concentration required for half-maximal activation of the ATPase is 4.7 +/- 0.2 muM for cardiac microsomes and 1.3 +/- 0.1 muM for skeletal microsomes at pH 6.8 and 0 degrees. The ATPase activities at saturating concentrations of ionized Ca2+ and pH 6.8, expressed as ATP hydrolysis per mg of protein, are 3 to 6 times lower for cardiac microsomes than for skeletal microsomes under a variety of conditions tested. The apparent Km value for MgATP at high concentrations in the presence of saturating concentrations of ionized Ca2+ is 0.18 +/- 0.03 ms at pH 6.8 and 25 degrees. The maximum velocity of ATPase activity under these conditions is 0.45 +/- 0.05 mumol per mg per min for cardiac microsomes and 1.60 +/- 0.05 mumol per mg per min for skeletal microsomes. The maximum steady state level of E approximately P for cardiac microsomes, 1.3 +/- 0.1 nmol per mg, is significantly less than the value of 4.9 +/- 0.2 nmol per mg for skeletal microsomes, so that the turnover number of the Ca2+-dependent ATPase of cardiac microsomes, calculated as the ratio of ATPase activity to the E approximately P level is similar to that of the skeletal ATPase. These findings indicate that the relatively slow rate of calcium transport by cardiac microsomes, whem compared to that of skeletal microsomes, reflects a lower density of calcium pumping sites and lower Ca2+ affinity for these sites, rather than a lower turnover rate. 相似文献
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The effects of an antiserum against the 53-kDa glycoprotein (GP-53) of the sarcoplasmic reticulum (SR) and of monoclonal antibodies against GP-53 on Ca2+ transport and ATP hydrolysis by SR of rabbit skeletal muscle have been investigated. Preincubation of SR with an antiserum against GP-53 resulted in decreased ATP-driven Ca2+ transport by the SR but had no effect on Ca2+-stimulated ATP hydrolysis. Preincubation of SR with preimmune serum had no significant effect on either Ca2+ transport or Ca2+-ATPase activity. The effect of anti-GP-53 serum was time and concentration dependent. Preincubation of SR with two monoclonal antibodies against GP-53 had no effect on Ca2+ transport or on Ca2+-stimulated ATP hydrolysis. However, preincubation of SR with either monoclonal antibody against GP-53 together with a monoclonal antibody against the Ca2+-ATPase (at levels which had little effect alone) resulted in markedly decreased rates of Ca2+ uptake and ATP hydrolysis. Preincubation of SR with anti-GP-53-serum or with monoclonal antibodies, under the same conditions that inhibited Ca2+ uptake, did not increase the passive permeability of the SR membrane to Ca2+, did not decrease the permeability of the SR to oxalate, and did not cause significant proteolysis of the Ca2+-ATPase. Our results are consistent with the interpretation that GP-53 may modulate the function of the Ca2+-ATPase of the SR membrane. 相似文献
8.
A J Murphy 《Biochemistry》1976,15(20):4492-4496
Modification of calcium-translocating sarcoplasmic reticulum membranes (SR) with 5,5'-dithiobis(2-nitrobenzoate) (Nbs2) reveals four classes (kinetic sets) of sulfhydryl groups. Of the 25 mol/1.5 X 10(5) G OF SR protein (i.e., containing 1 mol of ATPase protein) estimated in the presence of sodium dodecyl sulfate, 8 mol are unreactive, while 7, 8, and 2 mol display pseudo-first-order rate constants (k1) of 0.16, 0.68, and 8.3 min(-1), respectively (25 decrees C, pH 7.8, 4 MM Nbs2). Under these conditions, the Ca-ATPase activity is lost with k1 = 0.73 min(-1), whereas the Ca-independent ATPase activity is essentially unchanged. These results are little changed by the presence of Mg2+ or Ba2+ in the modification mixture, while Ca2+ or Sr2+ causes all 16-17 reactable sulfhydryls to be modified with k1 = 0.50 and 0.53 min(-1), respectively. The corresponding values for the loss of Ca-ATPase activity are 0.53 and 0.67 min(-1); this suggests that blocking of only one of the 16-17 SH groups inactivates the enzyme, i.e., that there is a single "essential" SH group. The midpoint of the transition between the Ca2+-free and Ca2+-modification patterns occurs at a free Ca2+ concentration of about 0.9 muM, implying that it is Ca2+ binding at the active sites (KD = 0.1 muM), rather than at the low-affinity nonspecific sites, that effects a conformation change in the ATPase protein (which contains greater than 90% of the cysteines). A calcium-induced conformation change is also suggested by increased ultraviolet absorbance spectrum of the purified ATPase protein upon calcium binding. If protein-lipid interaction is disrupted with deoxycholate or Triton X-100 (which does not destroy the Ca-ATPase activity and hence presumably leaves the tertiary structure of the ATPase protein largely intact), 95% of the sulfhydryls react with Nbs2 considerably faster; thus, at 2 mg/ml o- deoxycholate, 14 groups react with k1 greater than 20, 5 with k1 = 2.3, and 5 with k1 = 0.4 min(-1). These results suggest that the inaccessibility of SH groups in the absence of detergents is due to extensive interaction of the bilayer phospholipids with the ATPase protein. 相似文献
9.
The rates of calcium transport and Ca2+-dependent ATP hydrolysis by rabbit skeletal muscle sarcoplasmic reticulum were stimulated by monovalent cations. The rate of decomposition of phosphoprotein intermediate of the Ca2+-dependent ATPase of sarcoplasmic reticulum was also increased by these ions to an extent that is sufficient to account for the stimulation of calcium transport and Ca2+-dependent ATPase activity. The order of effectiveness of monovalent cations tested at saturating concentrations in increasing rate of phosphoprotein decomposition is: K+, Na+ greater than Rb+, NH4+ greater than Cs+ greater than Li+, choline+, Tris+. 相似文献
10.
Calcium transport by sarcoplasmic reticulum of vascular smooth muscle: I. MgATP-dependent and MgATP-independent calcium uptake. 总被引:1,自引:0,他引:1
M A Stout 《Journal of cellular physiology》1991,149(3):383-395
The components of 45calcium (Ca) uptake were studied in saponin skinned rat caudal artery. The steady-state Ca content increased when the free Ca concentration was varied from 10(-8) to 10(-4) M but was reduced by azide when the free Ca concentration exceeded 3.1 microM. The azide sensitivity and low affinity for Ca were consistent with functional mitochondria. The azide-insensitive component consisted of a small bound and a larger releasable Ca fraction. After skinning in Triton X-100, approximately 4 mumol Ca/kg wet tissue remained, which represented a tightly bound but slowly exchangeable Ca pool. The Ca content was independent of the free Ca concentration and MgATP, and it was not released with A-23187 or Ca. The Ca content of the larger fraction was a higher order function of the free Ca concentration and was released with A-23187, indicating it resided within a membrane-bounded structure. Ca uptake by the releasable fraction was increased by oxalate, MgATP, phosphocreatine, temperature, phosphate, and ruthenium red and represents Ca sequestered by the sarcoplasmic reticulum (SR) with little contribution from other Ca binding or storage sites. It is described by the coefficients Umax = 96.94 mumol/kg wet tissue, K1/2 = 0.75 microM, and Hill coefficient = 1.70. The SR in this preparation regulates cytosolic Ca concentrations under physiological conditions and can accumulate Ca by MgATP-dependent and MgATP-independent process. The larger, MgATP-dependent Ca uptake is described by the coefficients Umax = 72.87 mumol/kg wet tissue, K1/2 = 0.8 microM, and Hill coefficient = 2.09 and is consistent with Ca sequestered by the Ca-transport ATPase of smooth muscle SR. The smaller, MgATP-independent uptake is described by the coefficients Umax = 24.14 mumol/kg wet tissue, K1/2 = 0.56 microM, and Hill coefficient = 1.01 and represents Ca sequestered by an unidentified mechanism or by a subpopulation of SR. 相似文献
11.
Csala M Fulceri R Mandl J Benedetti A Bánhegyi G 《Biochemical and biophysical research communications》2001,287(3):696-700
We found that glutathione transport across endo/sarcoplasmic reticulum membranes correlates with the abundance of ryanodine receptor type 1 (RyR1). The transport was the fastest in muscle terminal cisternae, fast in muscle microsomes and slow in liver, heart, and brain microsomes. Glutathione influx could be inhibited by RyR1 blockers and the inhibitory effect was counteracted by RyR1 agonists. The effect of blockers was specific to glutathione, as the transport of other small molecules was not hindered. Therefore, the glutathione transport activity seems to be associated with RyR1 in sarcoplasmic reticulum. 相似文献
12.
Calcium phosphate precipitation in the sarcoplasmic reticulum reduces action potential-mediated Ca2+ release in mammalian skeletal muscle 总被引:1,自引:0,他引:1
During vigorous exercise, Pi concentration levels within the cytoplasm of fast-twitch muscle fibers may reach 30 mM. Cytoplasmic Pi may enter the sarcoplasmic reticulum (SR) and bind to Ca2+ to form a precipitate (CaPi), thus reducing the amount of releasable Ca2+. Using mechanically skinned rat fast-twitch muscle fibers, which retain the normal action potential-mediated Ca2+ release mechanism, we investigated the consequences of Pi exposure on normal excitation-contraction coupling. The total amount of Ca2+ released from the SR by a combined caffeine/low-Mg2+ concentration stimulus was reduced by 20%, and the initial rate of force development slowed after 2-min exposure to 30 mM Pi (with or without the presence creatine phosphate). Peak (50 Hz) tetanic force was also reduced (by 25% and 45% after 10 and 30 mM Pi exposure, respectively). Tetanic force responses produced after 30 mM Pi exposure were nearly identical to those observed in the same fiber after depletion of total SR Ca2+ by 35%. Ca2+ content assays revealed that the total amount of Ca2+ in the SR was not detectably changed by exposure to 30 mM Pi, indicating that Ca2+ had not leaked from the SR but instead formed a precipitate with the Pi, reducing the amount of available Ca2+ for rapid release. These results suggest that CaPi precipitation that occurs within the SR could contribute to the failure of Ca2+ release observed in the later stages of metabolic muscle fatigue. They also demonstrate that the total amount of Ca2+ stored in the SR cannot drop substantially below the normal endogenous level without reducing tetanic force responses. muscle fatigue; excitation-contraction coupling 相似文献
13.
A I Marzoev A G Maksina O A Azizova Iu A Vladimirov 《Biulleten' eksperimental'no? biologii i meditsiny》1980,89(4):410-412
The structure of the membranes of sarcoplasmic reticulum fragments (SRF) normally and in thyrotoxicosis was studied by the spin-label and spin-probe methods and by chemifluorescence. The curves of temperature dependence of the regularity parameter show a typical break for the spin probe at 20 degrees C shifted by 4 degrees C to sower temperatures for thyrotoxins. The same shift was observed with temperature dependence for the correlation period of the spin label covalently bound to the thiol groups of Ca2+ dependent ATPase of sarcoplasmic reticulum. The latent period of thyrotoxins was reduced and the chemifluorescence intensity increased. The results obtained suggest the occurrence of considerable changes in the structure of SRF membranes in thyrotoxicosis. 相似文献
14.
Synthesis of polyphosphoinositides has been studied in transverse (T-) tubule and sarcoplasmic reticulum (SR) membrane fractions of frog skeletal muscle, following 32P-labeling with [gamma-32P]ATP. Purified SR and T-tubule fractions respectively synthesize 9.4 +/- 0.8 and 71.9 +/- 9.8 pmol PtdInsP/mg per min, indicating nearly 8-fold higher activity of PtdIns kinase in the T-tubules than in the SR. The activity of this enzyme in both membrane systems is maximum at pH 7 and pCa 6. PtdInsP2 is synthesized from the endogenous PtdInsP, only in T-tubule membranes by the action of PtdInsP kinase. This lipid is the most intensely 32P-labeled phosphoinositide (181.7 +/- 9.2 pmol/mg per min) in these membranes. PtdIns kinase in the T-tubule and SR membranes, and PtdInsP kinase in the former are modulated by the free [Mg2+]. Loss of radiolabel from transiently maximal 32P-incorporation in polyphosphoinositides in T-tubule membranes, concomitant with a decrease in the ATP concentration in the incubation buffer, shows the occurrence of phosphoinositidases in these membranes. Under the conditions used, no such activities were evident in SR membranes. Compound 48/80, a mixture of condensation products of N-methyl-p-methoxyphenethylamine with formaldehyde, known to block phosphoinositidase C and phospholipase A2, causes a dose-dependent increase in the 32P-label of PtdInsP, in T-tubule membranes. The synthesis of lyso PtdInsP2, a deacylated form of PtdInsP2 which occurs in nearly equal quantities in both T-tubule and SR membranes, may result from a mechanism independent of phospholipase A2. 相似文献
15.
Calcium release from sarcoplasmic reticulum vesicles presumably derived from longitudinal tubules (LSR) and terminal cisternae (HSR) of rabbit skeletal muscle was investigated by dual wavelength spectrophotometry using the calcium-indicator antipyrylazo III. In 120 mM KCl, 5 mM MgCl2, 30 microM, CaCl2, 50 microM MgATP, 100 microM antipyrylazo III, 40 mM histidine (pH 6.8, 25 degrees C), LSR and HSR sequestered approx. 115 nmol calcium/mg, and then spontaneously released calcium. Analysis of ATP hydrolysis and phosphoenzyme level during LSR and HSR calcium sequestration indicated that this calcium release process was passive, occurring in the virtual absence of ATP and phosphoenzyme. Moreover, subsequent addition of ATP reinitiated the calcium sequestration-release sequence. Calcium release by HSR was more than 4-times faster than that by LSR. Analysis of the calcium release phase demonstrated a biexponential decay for both LSR (0.10 and 0.63 min-1) and HSR (0.26 and 1.65 min-1), suggestive of heterogeneity within each fraction. Replacement of 120 mM KCl with either 120 mM choline chloride, 240 mM sucrose, or H2O reduced maximal calcium sequestration by LSR, but had less effect on LSR calcium release rate constants. In the case of HSR, these changes in the ionic composition of the medium drastically reduced calcium release rate constants with little effect on calcium content. These marked differences between LSR and HSR are consistent with the hypothesis that the calcium permeability of the terminal cisternae is greater and more sensitive to the ionic environment than is that of the longitudinal tubules of sarcoplasmic reticulum. 相似文献
16.
Marie-Jeanne Loirat Brigitte Lucas-Heron Béatrice Ollivier Claude Leoty 《Bioscience reports》1988,8(4):369-378
Two Ca2+ sequestering proteins were studied in fast-twitch (EDL) and slow-twitch (soleus) muscle sarcoplasmic reticulum (SR) as a function of denervation time. Ca2+-ATPase activity measured in SR fractions of normal soleus represented 5% of that measure in SR fractions of normal EDL. Denervation caused a severe decrease in activity only in fast-twich muscle. Ca2+-ATPase and calsequestrin contents were affected differently by denervation. In EDL SR, Ca2+-ATPase content decreased progressively, whereas in soleus SR, no variation was observed. Calsequestrin showed a slight increase in both muscles as a function of denervation time correlated with increased45Ca-binding.These results indicate first that Ca2+-ATPase activity in EDL was under neural control, and that because of low Ca2+-ATPase activity and content in slow-twitch muscle no variation could be detected, and secondly that greater calsequestrin content might represent a relative increasing of heavy vesicles or decreasing of light vesicles as a function of denervation time in the whole SR fraction isolated in both types of muscles. 相似文献
17.
We investigated the effect of halothane on lipid and protein components of sarcoplasmic reticulum membranes isolated from pig trapezius muscle. We studied the relationships between the (Ca2(+)-Mg2+)-ATPase activity and the interaction of the anesthetic with lipid and protein moieties by means of EPR and fluorescence spectroscopic techniques. Our results clearly show that below 5 mumol per mg protein, halothane interacts mainly with the lipid components of the membrane. This interaction is shown to be localized in the central core of the phospholipid bilayer and to induce an increase of the membrane calcium permeability. The interaction with protein components only occurs at higher halothane concentrations and affects its conformational and functional states. These results are discussed with respect to new insights into diethylether-SR membrane interaction and to malignant hyperthermia syndrome in the pig. 相似文献
18.
Indu S. Ambudkar El-Sayed Abdallah Adil E. Shamoo 《Molecular and cellular biochemistry》1988,79(1):81-89
Summary The effects of various lysophospholipids on the calcium transport activity of sarcoplasmic reticulum (SR) from rabbit skeletal and canine cardiac muscles were examined. The lipids decreased calcium transport activity in both membrane types; the effectiveness being in the order lysoPC > lsyoPS, lysoPG > lysoPE. The maximum inhibition induced by lysoPC, lysoPG and lysoPS was greater than 85% of the normal Ca2+-transport rate. In cardiac SR lysoPE had a maximal inhibition of about 50%. Half maximal inhibition of calcium transport by lysoPC was achieved at 110 nmoles lysoPC/mg SR. At this concentration of lysoPC, the (Ca2+ + Mg2+)-ATPase and Ca2+-uptake activities were inhibited to the same extent (about 60%) in skeletal sarcoplasmic reticulum, while in cardiac sarcoplasmic reticulum, there was less than 20% inhibition of the Ca2+ + Mg2+-ATPase activity. Studies with EGTA-induced passive calcium efflux showed that up to 200 nmoles lysoPC/mg SR did not alter calcium permeability significantly in cardiac sarcoplasmic reticulum. In skeletal muscle membranes the lysophospholipid mediated decrease in calcium uptake correlated well with the increase in passive calcium efflux due to lysophosphatidylcholine. The difference in the lysophospholipid-induced effects on the sarcoplasmic reticulum from the two muscle types probably reflects variations in protein and other membrane components related to the respective calcium transport systems. 相似文献
19.
Z Mahrla 《Zhurnal evoliutsionno? biokhimii i fiziologii》1975,11(3):242-249
A comparative analysis of fatty acid composition of lipid components from skeletal muscle sarcolemma and sarcoplasmic reticulum membranes of rabbits and crayfishes Astacus fluviatilis and A. leptodactylus has been made by means of gas-liquid chromatography of methyl esters of fatty acids. There are slight differences between external and internal membranes in fatty acids composition of lipids in the same animal. Considerable differences were found, however, when lipids in corresponding membranes of different species were compared. There are more unsaturated fatty acids in the crayfish than in the rabbit membranes. The most expressive differences in fatty acid composition between the two animals concern the content of linoleic acid type. Polyunsaturated acids in the crayfish are mostly of a 3 type and those in the rabbit of a 6 type. During biochemical evolution the following changes seem to take place, a decrease in the amount of unsaturated fatty acids as well as of polyunsaturated ones of a 3 type and an increase of those of the 6 type. These changes very probably reflect the transition from an aquatic to a terrestrial habitat. 相似文献
20.
O M Rozhmanova G Ia Dubur V V Kastron L N Stel'makh S G Voronina 《Biokhimii?a (Moscow, Russia)》1988,53(10):1612-1618
Using the radioisotope method, the Ca2+ transport through proteoliposomes was investigated. The proteoliposomes originating from total brain lipids and skeletal muscle T-system membranes of the rabbit were shown to possess a Ca2+ permeability which can be stimulated by 1.4-dihydroxypyridine derivatives (10(-9)-10(-7) M). Verapamil and Cd2+ (10(-5) M and 10(-3) M, respectively) inhibit the Ca2+ permeability of proteoliposomes stimulated by dihydroxypyridine derivatives. The activating effect of the latter depends on the microviscosity of the proteoliposome lipid bilayer. An addition of cholesterol to brain phospholipids at a ratio of 1:5 increases the stimulating effect of dihydroxypyridine by 50%. 相似文献