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1.
T Kitazawa 《Journal of biochemistry》1976,80(5):1129-1147
It was investigated whether mitochondria play a significant role in the physiological regulation of the contractile process by Ca2+ in cardiac muscle in comparison with the sarcoplasmic reticulum (SR). Ca uptake activities of chicken cardiac SR and rabbit cardiac mitochondria were measured by means of centrifugation, dual-wave-length spectrophotometric and Millipore filtration methods. The maximum Ca uptake capacity of cardiac SR was usually 50-60 nmoles/mg protein and the apparent binding constant was 2.0 X 10(6) M-1. The apparent Ca-binding constant of cardiac mitochondria under limited loading conditions was 2.4 X 10(5) M-1 at pH 7.4 and 5.9 X 10(4) M-1 at pH 6.8. In the presence of 100 muM Ca2+ at 28-29 degrees, the estimated initial rate of Ca uptake of cardiac SR ranged from 20 to 30 nmoles Ca/mg-sec, while that of mitochondria was 4.6 nmoles Ca/mg-sec under limited loading conditions at pH 7.4 and 0.64 nmoles Ca/mg-sec under massive loading conditions at pH 6.8, which was much closer to physiological conditions. In the presence of low Ca2+ concentrations, the initial rate of Ca uptake of cardiac SR was 0.5 nmoles Ca/mg-sec at 3.5 X 10(-7) M Ca2+ and that of mitochondria under massive loading conditions at 1 X 10(-6) M Ca2+ was 0.02 nmoles Ca/mg-sec at pH 7.4 and 0.004 nmoles Ca/mg-sec at pH 6.8. The Ca uptake activities were also examined using glycerol-extracted cardiac muscle fibers. Cardiac SR, 1.7 mg/ml, reduced the tension of maximally contracted cardiac muscle fibers to a level corresponding to about 30% of maximum tension, but in the presence of 14.3 mg/ml of mitochondria the maximum tensions of both skeletal muscle and cardiac muscle fibers were maintained for at least 3 min. From these results the time course of relaxation of cardiac muscle induced by cardiac SR or mitochondria was calculated. It was concluded that, in the physiological contraction of cardiac muscle, the SR plays a major role in controlling intracellular Ca2+ movement; the Ca uptake of mitochondria is relatively insignificant. When the cardiac muscle contracts maximally, SR alone cannot relax the cardiac muscle without the aid of other Ca removing system. 相似文献
2.
Steady-state properties of calcium binding to parvalbumins from bullfrog skeletal muscle: effects of Mg2+, pH, ionic strength, and temperature 总被引:1,自引:0,他引:1
To improve our understanding of the physiological roles of parvalbumins, PA-1 (pI 4.78) and PA-2 (pI 4.97) parvalbumins were prepared from bullfrog skeletal muscle and their calcium binding properties were examined in a medium of constant ionic strength (I = 0.106, pH 6.80, at 20 degrees C) containing various concentrations of Mg2+ by using a metallo-indicator, tetramethylmurexide. Apparent binding constants for Ca2+ in the presence of Mg2+ changed in the manner expected if Ca2+ and Mg2+ compete for two independent homogeneous binding sites. The following values were obtained: for PA-1, KCa = 1 X 10(7) M-1, KMg = 900 M-1; for PA-2, KCa = 6 X 10(6) M-1, KMg = 830 M-1 (I = 0.106, pH 6.80, at 20 degrees C). The apparent binding constants are strongly dependent on temperature: at 10 degrees C for PA-1, KCa = 2 X 10(8) M-1, KMg = 10(4) M-1; for PA-2, KCa = 5 X 10(7) M-1, KMg = 5 X 10(3) M-1 (I = 0.106, pH 6.80). The dependence of the affinities for Ca2+ on ionic strength is similar to or less than that of GEDTA (EGTA). The affinities for Ca2+ and Mg2+ of parvalbumins are unchanged between pH 6.5 and 7.2. 相似文献
3.
The effects of pH, aluminium concentration and temperature on the embryonic development of the European common frog, Rana temporaria 总被引:1,自引:0,他引:1
The individual and combined effects of pH, aluminium concentration and temperature, on the development of common frog ( Rana temporaria ) embryos from an upland area of northern England, were investigated in a controlled laboratory study. There was strong evidence to suggest that embryonic survival was lower at pH 4.5 compared with pH 6.0. At pH 4.5, embryonic survival was reduced at the highest aluminium concentrations. There was no strong evidence for a reduction in embryonic survival at lower aluminium concentrations.
Gastrulation and hatching appeared to be the most sensitive stages to both pH and aluminium concentration.
There was strong evidence to suggest that temperature-shocked embryos had reduced survival when compared with those kept at a constant temperature, particularly at pH 4.5.
There was substantial variation in the survival of embryos from different clutches in the same pond with respect to low pH, aluminium concentration and temperatures. Thus R. temporaria has a wide genetic base from which to tolerate environmental changes. It is suggested, however, that the lethal and sublethal effects of low pH, high aluminium concentration and low temperature, could lead to a decrease in recruitment to the adult populations of R. temporaria in upland, northern England. 相似文献
Gastrulation and hatching appeared to be the most sensitive stages to both pH and aluminium concentration.
There was strong evidence to suggest that temperature-shocked embryos had reduced survival when compared with those kept at a constant temperature, particularly at pH 4.5.
There was substantial variation in the survival of embryos from different clutches in the same pond with respect to low pH, aluminium concentration and temperatures. Thus R. temporaria has a wide genetic base from which to tolerate environmental changes. It is suggested, however, that the lethal and sublethal effects of low pH, high aluminium concentration and low temperature, could lead to a decrease in recruitment to the adult populations of R. temporaria in upland, northern England. 相似文献
4.
H Lorkovi? 《Canadian journal of physiology and pharmacology》1979,57(2):217-219
Membrane potential changes and mechanical tension provoked by 20 or 80 mM K at pH 8.0 or 4.5 were measured in muscle fiber bundles from Rana temporaria and Rana esculenta. The bathing solution contained Cl as the main anion. Low pH increased the depolarizing efficiency of high K to about the same degree in muscles of both species of frogs. The tension-[K] relationship was shifted toward lower [K] in muscles from R. temporaria and toward higher [K] in muscles from R. esculenta. It was concluded that the depolarization-contraction coupling was insensitive to pH changes in the former but not in the latter frog species. 相似文献
5.
O Poupa K B Helle M Lomsky 《Comparative biochemistry and physiology. A, Comparative physiology》1985,81(4):801-805
In seven poikilotherm species, i.e. cyclostome, Myxine glutinosa; teleosts, Gadus morrhua; Platichtys flesus; amphibia, R. pipiens; R. temporaria; Xenopus laevis; reptiles, Vipera berus. Ca paradox could not be obtained in ventricular strips at 8-12 degrees C. In similar preparations from human atria at 30 degrees C and 37 degrees, Ca paradox was obtained similarly as in other homoiotherms. In homoiotherms, lowered temperature (below 30 degrees C) has a protective effect. Viper myocardium did not display Ca paradox at 12, 22 or 32 degrees C, whereas human myocardium displayed Ca paradox even at 30 C. It is postulated that the absence of Ca paradox in poikilotherms is not due to the low temperature. In hagfish (Myxine glutinosa) the absence of Ca paradox seems to be due to the specific conditions concerning Ca exchange in the myocardial cell (large glycocalyx). 相似文献
6.
Tonic contraction and the control of relaxation in a chemically skinned molluscan smooth muscle 总被引:1,自引:0,他引:1
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F Cornelius 《The Journal of general physiology》1982,79(5):821-834
The same functional states that characterize the living anterior byssus retractor muscle (ABRM) from Mytilus edulis can be initiated in the saponin-treated (chemically skinned) muscle preparation under controlled biochemical conditions. A tonic contraction was induced if the concentration of free Ca2+ was above approximately 10(7) M in the presence of Mg2+ and ATP. Maximum tension development was achieved at a Ca2+ concentration of approximately 10(4) M. Within these Ca2+ concentrations tension was always associated with the presence of 'active state," as indicated by a high recovery of tension after a quick release in muscle length. Tonic tension, and the associated active state was maintained for hours during these conditions irrespective of variations in both ionic strength and pH. Reduction of the Ca2+ concentration to below threshold for tension initiation during a tonic contraction immediately switched off the active state and relaxation of the muscle preparation resulted. However, the rate of relaxation was extremely low, leaving a substantial fraction of tension in the absence of active state. Both 5-hydroxytryptamine (5-HT) and cAMP accelerated this slow relaxation in the absence of Ca2+. Thus, this state was considered equivalent to the 'catch state" in the living ABRM. In the presence of Ca2+ concentrations above 10(7) M, cAMP did not affect either the maximum tension developed or the Ca2+ sensitivity of the chemically skinned muscle preparation. 相似文献
7.
H Shi P H Hamm R G Lawler D C Jackson 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》1999,122(2):173-180
We compared responses of turtle heart at 20 degrees C to an anoxic lactic acidosis solution (LA) containing 35 mM lactic acid in an otherwise normal turtle Ringers equilibrated with 3% CO2/97% N2 at pH 7.0) to a solution simulating in vivo anoxic acidosis (VA), with elevated concentrations of lactate, Ca2+, Mg2+, and K+, and decreased Cl-, equilibrated with 10.8% CO2/89.2% N2 at pH 7.0. We examined mechanical properties on cardiac muscle strips and determined intracellular pH (pHi) and high energy phosphates on perfused hearts using 31P-NMR. Maximum active force (Fmax) and the maximum rate of force development (dF/dtmax) of muscle strips were significantly higher during VA than during LA superfusion. An elevation of Ca2+ alone (to 6 mM) in LA significantly increased both Fmax and dF/dtmax but the effects diminished toward the end of the exposure; however, hypercapnic anoxic lactic acidosis (addition of 20 mM HCO3- to LA, equilibrated with 10.8% CO2/89.2% N2, pH 7.0) did not significantly affect Fmax or dF/dtmax. During VA perfusion, pHi (6.73 +/- 0.01) was significantly higher than that during LA perfusion (pHi 6.69 +/- 0.013), but the difference is probably too small to have physiological significance. ATP, creatine phosphate, and inorganic phosphate were not significantly different in the two anoxic solutions. We conclude that the reduction of cardiac mechanical function in vivo is minimized by the integrated effects of changes of ionic concentrations, but the observed changes in Ca2+ and pHi cannot fully explain the effect. 相似文献
8.
Sensitivity of cultured chick myotubes to alkaline earth metal ions was investigated by recording contractile isometric tension through a semiconductor transducer. The myotubes were obtained by culturing myoblasts of chick embryo breast muscles, and skinned chemically before physiological experiments. Contractions developed in response to Ca2+ in a bathing medium higher than 3 x 10(-7) M and reached maximum at 1 x 10(-5) M. Sr2+ was less effective than Ca2+; the threshold concentration was 1 x 10(-5) M and the tension reached maximum at 1 x 10(-3) M. Ba2+ was the least effective among the three alkaline earth metal ions; only one fifth of the Ca(2+)-induced maximum tension was attained at 1 x 10(-3) M. The sensitivity was similar to that of the mature pectoral muscle fiber, a fast twitch muscle fiber, rather than that of the anterior latissimus dorsi, a slow tonic muscle fiber. The sensitivity was shown to be dependent on its troponin C by replacing it with troponin C from the mature pectoral or cardiac muscle. This indicates that TnC of a fast-muscle type is expressed in the cultured chick myotube as in the mature pectoral muscle. The contractile apparatus was thus shown to be well developed in the cultured myotube with characteristics similar to the mature fast twitch muscle fiber. 相似文献
9.
The concentration of intracellular free calcium ions in the follicle wall cells and in the follicle cells of Rana temporaria in Ringer solution is 150 +/- 10 and in the follicle wall cells of Xenopus laevis, 220 +/- 10 nM. In a chloride-free saline, its concentration in the same cells is 2.5-3 times that in Ringer solution. Voltage-dependent Ca(2+)-channel blockers diltiazem and verapamil (100 microM) reduce the level of intracellular free calcium ions in R. temporaria follicle wall cells cultivated in a chloride-free saline to 170 +/- 20 nM, which practically does not differ from the level in Ringer solution. Inhibitors (100 microM) decrease the rate of "spontaneous" maturation of R. temporaria follicle-enclosed oocytes both in chloride-free and Ringer solutions. It was concluded that an increased level of intracellular free calcium ions in the follicle cells, among other factors, may determine the stimulating effect of the medium (Ringer or chloride-free solution) on "spontaneous" maturation of follicle-enclosed amphibian oocytes. Voltage-dependent calcium channels appear to be involved in Ca2+ influx into the cells. 相似文献
10.
A protein inhibitor of the Ca2+-dependent proteinase has been purified from bovine cardiac muscle by using the following steps in succession: salting out 17,600 X gmax supernatants from muscle homogenates in 50 mM Tris acetate, pH 7.5, 4 mM EDTA between 25 and 65% ammonium sulfate saturation; eluting between 25 and 120 mM KCl from a DEAE-cellulose column at pH 7.5; salting out between 30 and 60% ammonium sulfate saturation; Ultrogel-22 gel permeation chromatography at pH 7.5; heating to 80 degrees C followed by immediate cooling to 0 degree C; 6% agarose gel permeation chromatography in 4 M urea, pH 7.5; and elution from a phenyl-Sepharose hydrophobic column between 0.7 and 0.5 M ammonium sulfate. Approximately 1.16-1.69 mg of purified Ca2+-dependent proteinase inhibitor are obtained from 1 kg of bovine cardiac muscle, fresh weight. Bovine cardiac Ca2+-dependent proteinase inhibitor has an Mr of 115,000 as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a pI of 4.85-4.95, very little alpha-helical structure, a very low specific absorbance of 1.647 (A1% 280), and very low contents of histidine, tryptophan, phenylalanine, and tyrosine. Bovine cardiac Ca2+-dependent proteinase inhibitor probably contains a single polypeptide chain in nondenaturing solvents. One 115-kDa inhibitor polypeptide inactivates 10 110-kDa millimolar Ca2+-requiring proteinase (millimolar Ca2+-dependent proteinase) molecules in assays of purified proteins. Inhibition of millimolar proteinase by the proteinase inhibitor did not change in the pH range 6.2-8.6. The inhibitor requires Ca2+ to bind to millimolar Ca2+-dependent proteinase. The Ca2+ concentration required for one-half-maximum binding of millimolar Ca2+-dependent proteinase to the inhibitor was 0.53 mM, compared with a Ca2+ concentration of 0.92 mM required for one-half maximum activity of millimolar Ca2+-dependent proteinase in the absence of the proteinase inhibitor. Unless millimolar Ca2+-dependent proteinase is located subcellularly in a different place than the proteinase inhibitor or unless the proteinase/inhibitor interaction is regulated, millimolar proteinase could never be active in situ. 相似文献
11.
The types and the distribution of muscle fibres were analysed and compared in the tails of Xenopus laevis and Rana temporaria tadpoles. The filter feeding tadpoles of X. laevis were found to have both white muscle fibres adjacent to the notochord used for normal locomotory swimming and a superficial layer of small red fibres. The red fibres are probably used for the continuous flickering movement of the tail associated with the maintenance of the mid-water filter feeding position. R. temporaria, a grazing detritus feeding tadpole, was found to have only white muscle fibres used for normal locomotory swimming. Smaller superficial fibres were not red fibres but were thought to be immature white fibres. 相似文献
12.
The absorption spectrum of arsenazo III in media containing K+, Mg2+ and Ca2+ is sharply influenced by pH in the range of 7.5--5.0. The effect of pH is particularly pronounced in the wavelength range 532--602 nm due to the large pH dependence of the dissociation constant of Mg-arsenazo III complex. Therefore absorption changes at these wavelengths during muscle contraction cannot be used as reliable indicators of free ionized Ca2+ concentration in the cell. The effect of pH is less pronounced, but still noticeable at the wavelength pairs 575--650 or 660--685 nm. Multiple layers of muscle cells grown on polystyrene coils permit measurement of absorption changes of arsenazo III, introduced into the cells, by equilibration with 0.5 mM arsenazo III under routine culture conditions. The absorbance changes recorded at 660--685 nm are probably related to changes in intracellular free Ca2+ concentration. 相似文献
13.
The motility of demembranated bull sperm was found to be governed by the concentrations of cyclic adenosine 3', 5'-monophosphate (cAMP) and Ca2+ at low pH (6.6-7.1), and was less sensitive to these variables at higher pH (7.4-7.8). Although motility was generally found to increase with increasing pH in the range from 6.6 to 7.8, the addition of exogenous cAMP markedly and selectively improved the motility at the lower end of the range (pH 6.6-7.1). In the presence of 10 microM cAMP, low Ca2+ (8.0 X 10(-8) M), and a high concentration of Mg-adenosine 5'-triphosphate (ATP, 8 mM), demembranated sperm at pH 6.8 and 7.1 exhibited swimming similar to that of live ejaculated sperm. At a free Ca2+ concentration of 4.4 X 10(-5) M, the motility was rapidly inhibited at pH 6.8-7.1, whereas at pH 7.4-7.8, the activity was not greatly affected. Since calcium is known to antagonize the cAMP pathway by activating Ca2+-dependent phosphodiesterase and Ca2+-dependent phosphatase, this further supports the idea that cAMP-dependent activation is crucial for motility at low pH. Our results demonstrate that the flagellar axoneme can function normally at relatively acidic pH, and produce vigorous swimming at high levels of ATP. The ATP content of live sperm was measured and found to be high enough (approximately 8 mM) to support the vigorous motility seen at pH 6.6-7.1 in the models.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
14.
Maturation of Rana temporaria and Xenopus laevis oocytes was induced by solutions containing Mn2+ and Co2+ ions. Completion of oocyte maturation was estimated by the following criteria: (1) appearance of the maturation promoting factor (MPF) in the oocyte cytoplasm and (2) oocyte capacity to activation and formation of male pronuclei from the injected sperm nuclei. X. laevis oocytes matured under the effect of Co2+ ions were shown to contain MPF. Oocytes of both species matured under the effect of either ions could not be activated by pricking with a needle and injected sperm nuclei didn't transform into pronuclei. R. temporaria oocytes matured under the effect of ions in late spring, when natural spawning takes place, showed spontaneous activation. 相似文献
15.
Vascular smooth muscle contracts on increases of extracellular pH (pH(o)) and relaxes on pH(o) decreases possibly resulting from changes in transsarcolemmal Ca(2+) influx. Therefore, we studied store-operated Ca(2+) entry (SOCE; i.e. capacitative Ca(2+) entry (CCE)) during acidification (pH(o)=6.5) and alkalinization (pH(o)=8.0) in isolated porcine coronary smooth muscle cells (SMCs) by monitoring cytoplasmic Ca(2+) ([Ca(2+)](i)) and divalent cation entry (Mn(2+) quench) with fura-2/AM-fluorometry. Additionally, we evaluated the contribution of SOCE to pH(o)-dependent changes in isometric tension of porcine coronary smooth muscle strips. SOCE elicited in SMCs by the SERCA inhibitor BHQ was strongly modulated by pH(o) showing a decrease upon acidification and vice versa an increase upon alkalinization. BHQ-mediated tension of smooth muscle strips also revealed strong pH(o) dependence. In contrast, L-VOC-dependent tension ([K(+)](o)=20 and 40 mmol l(-1)) was remarkably less affected by pH(o) changes. Moreover, refilling of depleted Ca(2+) stores after repeated M(3)-cholinergic receptor stimulation could be almost completely inhibited by SKF 96365 and was markedly reduced by acidification and considerably enhanced by alkalinization pointing to a major role of SOCE in refilling. We conclude that vascular tone particularly responds to alterations in pH(o) whenever SOCE substantially contributes to the amount of activator Ca(2+) for contraction. 相似文献
16.
The effects of zero extracellular Ca2+ on the contractility of rat diaphragmatic strips in vitro were studied in conjunction with various pharmacological agents known to influence the intracellular Ca2+ concentration: the Na+ ionophore, monensin, and the Na(+)-K+ pump inhibitor, ouabain, which enhance [Ca2+]i, caffeine, which induces Ca2+ release from the sarcoplasmic reticulum (SR), and ryanodine, which prevents Ca2+ retention by the SR. The effect of increasing [Ca2+]i on diaphragmatic contraction was assessed by comparing contractions induced by 120 mM K+ in the small muscle strips before and after the addition of ouabain or monensin. Monensin (20 microM) and ouabain (1-100 microM) augmented contractions up to threefold. Treatment of diaphragm strips with 3 nM ryanodine increased baseline tension 360% above the original resting tension but only if the diaphragm was electrically stimulated concurrently; 100 microM ryanodine induced contracture in quiescent tissue. High K+ contractures were of greater magnitude in the presence of ryanodine compared with control, and relaxation time was prolonged by greater than 200%. Ca(2+)-free conditions ameliorated these actions of ryanodine. Ryanodine reduced contractions induced by 10 mM caffeine and nearly abolished them in Ca(2+)-free solution. The data demonstrate that extracellular Ca2+ is important in certain types of contractile responses of the diaphragm and suggest that the processes necessary to utilize extracellular Ca2+ are present in the diaphragm. 相似文献
17.
The regulation of intracellular pH (pHi) is important in controlling muscle contraction. In these experiments, a spectrophotometric method of determining pHi was developed, and the method was then used to study muscle pHi regulation during CO2-induced changes in extracellular pH (pHb). Studies were performed in vitro on 27 diaphragm muscle strips obtained from adult hamsters. pHi was measured from the ratio of the absorbances of the acid (lambda = 530 nm) and alkaline (lambda = 460 nm) forms of a vital dye, neutral red, using the unstained diaphragm spectrum as a reference blank. A standard neutral red calibration curve constructed from eight diaphragm muscle homogenates indicated that the absorbance ratio was highly linear, with pH over the range 6.00-8.00. In intact muscle strips gassed with 95% O2-5% CO2, pHb was 7.45 +/- 0.03 (SE) and pHi was 7.00 +/- 0.01 (SE). When the muscle was aerated with CO2 concentrations from 3 to 30%, pHb and pHi changed rapidly and reached a steady state in 10-15 min. However, when pHb ranged from 6.80-7.80, pHi changed little from the value observed when pHb was 7.40. When pHb was less than 6.80 or greater than 7.80, changes in pHi and pHb were quantitatively similar. The results suggest that, in the isolated diaphragm, overall pHi is stable and effectively buffered over a wide range of CO2-induced changes in buffer solution pH. 相似文献
18.
Effects of pH, temperature, and calcium concentration on the stoichiometry of the calcium pump of sarcoplasmic reticulum 总被引:2,自引:0,他引:2
Coupling of Ca2+ transport to ATP hydrolysis by isolated skeletal muscle sarcoplasmic reticulum vesicles has been investigated by means of ATP pulse methods. The stoichiometric amounts of Ca2+ transported per pulse of ATP were measured by Ca2+-stat methods, using either a Ca2+ electrode or arsenazo III as end point detectors, or by means of 45CaCl2. Maximum coupling ratios (Ca2+/ATP), of 1.82 +/- 0.13 occurred at pH 6.8, 25 degrees C, and in the presence of saturating Ca2+ concentrations. Ca2+/ATP values decreased at alkaline pH, with an apparent pK alpha of 7.9. The coupling ratio was unaltered between 6 and 30 degrees C, but decreased to 0.4 at 42 degrees C. Uncoupling by alkaline pH and high temperatures was reversible. The coupling process was Ca2+-dependent, with a K0.5 value for Ca2+ of 0.12 microM and a Hill coefficient of 2.0. Ca2+ ions, which were transported into vesicles under conditions resulting in low coupling ratios, were retained as the calcium oxalate precipitate, following complete hydrolysis of substrate. Passive Ca2+ efflux and Ca2+ exchange, were independent of pH. The observed variations in Ca2+/ATP ratio cannot readily be explained on the basis of a pump-leak model. Rather, the Ca2+-ATPase appears to be capable of pumping Ca2+ ions, under physiological conditions, with variable stoichiometry that is dependent upon its thermodynamic loading. 相似文献
19.
Irreversible thiophosphorylation and activation of tension in functionally skinned rabbit ileum strips by [35S]ATP gamma S. 总被引:4,自引:0,他引:4
Rabbit ileum strips were functionally skinned by exposure to staphylococcal alpha-toxin. Incubation of the strips in the ATP analog ATP gamma S or [35S]ATP gamma S in the presence of Ca2+ (but not in the absence of Ca2+) resulted in a maximal Ca2+-insensitive activated tension that persisted following removal of Ca2+. Correlated with this tension was 35S-labeling of the 20,000-dalton myosin light chain, LC20, that persisted even after removal of Ca2+. Tension in these strips partially relaxed when exposed to ATP (alpha,beta-methylene). In contrast, alpha-toxin-treated strips exposed to ATP or [gamma-32P]ATP showed Ca2+-sensitive, reversible activated tension and reversible 32P-labeling of the LC20. These results are consistent with a currently proposed model of Ca2+ control of smooth muscle contraction involving a myosin light chain kinase-phosphatase system. 相似文献
20.
Evidence for a calcium-sensitive factor which alters the alkaline pH sensitivity of sarcoplasmic reticulum calcium transport 总被引:1,自引:0,他引:1
C A Tate A Chu J McMillin-Wood W B Van Winkle M L Entman 《The Journal of biological chemistry》1981,256(6):2934-2939
Oxalase-supported, ATP-dependent Ca2+ uptake by cardiac and skeletal muscle sarcoplasmic reticulum (SR) exhibits a pH profile with the maximal rate of Ca2+ uptake at pH 6.6-6.8 and marked inhibition (90-95%) at pH 7.4-7.6, a point at which Ca2+-dependent ATPase activity is optimal. These observations are noted when the SR is first preincubated in media containing no added Ca2+. This alkaline pH inhibition is not caused by an irreversible perturbation since the Ca2+ uptake rate is fully restored by changing the alkaline pH preincubation medium to pH 6.8. When SR is preincubated with added Ca2+, Ca2+ uptake at alkaline pH (7.4-7.6) is only inhibited by 10-30%. Ca2+ uptake at pH 6.8 is the same regardless of preincubation conditions. A depressed oxalate permeability is not a factor in the observed alkaline pH inhibition of Ca2+ uptake. At alkaline pH, the relationship between the preincubation Ca2+ concentration and the rate of Ca2+ uptake is hyperbolic; the half-maximal free Ca2+ concentration for stabilization of Ca2+ uptake is 8-15 microM with a Vmax equal to the velocity at the optimal pH. The Hill coefficient is 1.0, implying a single class of Ca2+-requiring sites for stabilization at alkaline pH. In contrast to its effect on Ca2+ uptake, the presence of Ca2+ during preincubation does not alter the pH sensitivity of Ca2+-dependent ATPase activity. Thus, the presence of Ca2+ during preincubation may stabilize a state of the CaATPase, conducive to the coupling of net Ca2+ translocation to Ca2+-dependent ATPase activity, which is ordinarily opposed by alkaline pH. The data suggest a single class of Ca2+-requiring sites which favors this coupled state. 相似文献