Preferential support of Ca 2+ uptake in smooth muscle plasma membrane vesicles by an endogenous glycolytic cascade

1989 ◽  
Vol 3 (11) ◽  
pp. 2298-2301 ◽  
Author(s):  
Richard J. Paul ◽  
Christopher D. Hardin ◽  
Luc Raeymaekers ◽  
Frank Wuytack ◽  
Rik Casteels
Keyword(s):  
Smooth Muscle ◽  
Plasma Membrane ◽  
Membrane Vesicles ◽  
Plasma Membrane Vesicles ◽  
Muscle Plasma Membrane

The effects of acetylcholine and atropine on the 22Na+ permeability of intestinal smooth muscle vesicles

1978 ◽  
Vol 56 (6) ◽  
pp. 921-925
Author(s):  
L. Spero
Keyword(s):  
Smooth Muscle ◽  
Plasma Membrane ◽  
Membrane Vesicles ◽  
Intestinal Smooth Muscle ◽  
Erythrocyte Ghosts ◽  
Muscarinic Agonists ◽  
Plasma Membrane Vesicles ◽  
Muscle Plasma Membrane ◽  
Whole Muscle ◽  
Kinetics Of

A technique is described which has enabled us to measure changes in 22Na+ efflux from smooth muscle plasma membrane vesicles. The resting 22Na+ efflux from these sealed vesicles showed a concentration-dependent increase in response to acetylcholine and other muscarinic agonists, in similar concentrations to those which increased 42K+ efflux in whole muscle. The kinetics of this efflux were complex and could not be described by less than three exponential processes. The response to agonists has, therefore, been characterized by measurement of the half-life of 22Na+ efflux (t1/2). The acetylcholine effect was inhibited by atropine, but unlike the situation in the whole muscle, this inhibition was noncompetitive. Tubocuraine (a nicotinic antagonist) had no effect on this acetylcholine response. Atropine has no effect by itself on the resting 22Na+ efflux, neither did tetrodotoxin or ouabain. 22Na+ efflux from erythrocyte ghosts and liposomes, prepared from lipid extracts of the smooth muscle plasma membrane, was not modified by acetylcholine or atropine.

scholarly journals Comparison of endogenous and exogenous sources of ATP in fueling Ca2+ uptake in smooth muscle plasma membrane vesicles.

1992 ◽  
Vol 99 (1) ◽  
pp. 21-40 ◽  
Author(s):  
C D Hardin ◽  
L Raeymaekers ◽  
R J Paul
Keyword(s):  
Smooth Muscle ◽  
Plasma Membrane ◽  
Calcium Uptake ◽  
Membrane Vesicles ◽  
Glycolytic Enzyme ◽  
Calcium Pump ◽  
Bulk Solution ◽  
Plasma Membrane Vesicles ◽  
Atp Production ◽  
Muscle Plasma Membrane

A smooth muscle plasma membrane vesicular fraction (PMV) purified for the (Ca2+/Mg2+)-ATPase has endogenous glycolytic enzyme activity. In the presence of glycolytic substrate (fructose 1,6-diphosphate) and cofactors, PMV produced ATP and lactate and supported calcium uptake. The endogenous glycolytic cascade supports calcium uptake independent of bath [ATP]. A 10-fold dilution of PMV, with the resultant 10-fold dilution of glycolytically produced bath [ATP] did not change glycolytically fueled calcium uptake (nanomoles per milligram protein). Furthermore, the calcium uptake fueled by the endogenous glycolytic cascade persisted in the presence of a hexokinase-based ATP trap which eliminated calcium uptake fueled by exogenously added ATP. Thus, it appears that the endogenous glycolytic cascade fuels calcium uptake in PMV via a membrane-associated pool of ATP and not via an exchange of ATP with the bulk solution. To determine whether ATP produced endogenously was utilized preferentially by the calcium pump, the ATP production rates of the endogenous creatine kinase and pyruvate kinase were matched to that of glycolysis and the calcium uptake fueled by the endogenous sources was compared with that fueled by exogenous ATP added at the same rate. The rate of calcium uptake fueled by endogenous sources of ATP was approximately twice that supported by exogenously added ATP, indicating that the calcium pump preferentially utilizes ATP produced by membrane-bound enzymes.

scholarly journals Characterization of L-carnitine transport into rat skeletal muscle plasma membrane vesicles

2000 ◽  
Vol 267 (7) ◽  
pp. 1985-1994 ◽  
Author(s):  
Simona Berardi ◽  
Bruno Stieger ◽  
Bruno Hagenbuch ◽  
Ernesto Carafoli ◽  
Stephan Krähenbühl
Keyword(s):  
Skeletal Muscle ◽  
Plasma Membrane ◽  
Membrane Vesicles ◽  
Rat Skeletal Muscle ◽  
Plasma Membrane Vesicles ◽  
Muscle Plasma Membrane ◽  
Carnitine Transport

Effect of disulphonic stilbenes on Ca2+ transport in smooth muscle plasma membranes

1984 ◽  
Vol 62 (9) ◽  
pp. 1233-1238 ◽  
Author(s):  
P. K. Rangachari ◽  
A. K. Grover ◽  
E. E. Daniel
Keyword(s):  
Smooth Muscle ◽  
Plasma Membrane ◽  
Plasma Membranes ◽  
Circular Muscle ◽  
Membrane Vesicles ◽  
Plasma Membrane Vesicles ◽  
Disulphonic Acid

We studied the effects of two disulphonic stilbenes, 4′,4′-diisothiocyano-2,2′-stilbene disulphonic acid (DIDS) and 4-acetamido-4′-isothiocyano-2,2′-stilbene disulphonic acid (SITS), on Ca2+ transport by plasma membrane vesicles from the circular muscle of the dog stomach. Both compounds inhibited ATP-dependent Ca2+ uptake and reduce the leak from loaded vesicles. The inhibition produced could not be significantly reduced by either permeant anions or by increasing the level of free Ca2+. The effects of DIDS could be rendered irreversible by incubating the membranes with this agent at 37 °C.

Characterization of Ca2+ uptake in plasma membrane vesicles isolated from guinea pig ileum smooth muscle

Cell Calcium ◽  
1987 ◽  
Vol 8 (1) ◽  
pp. 65-77 ◽  
Author(s):  
Ram V. Sharma ◽  
Carol A. Butters ◽  
James P. McEldoon ◽  
Ramesh C. Bhalla
Keyword(s):  
Smooth Muscle ◽  
Plasma Membrane ◽  
Guinea Pig ◽  
Membrane Vesicles ◽  
Guinea Pig Ileum ◽  
Plasma Membrane Vesicles

scholarly journals Isolation of a plasma-membrane fraction from gastric smooth muscle. Comparison of the calcium uptake with that in endoplasmic reticulum

1983 ◽  
Vol 210 (2) ◽  
pp. 315-322 ◽  
Author(s):  
L Raeymaekers ◽  
F Wuytack ◽  
J Eggermont ◽  
G De Schutter ◽  
R Casteels
Keyword(s):  
Endoplasmic Reticulum ◽  
Smooth Muscle ◽  
Plasma Membrane ◽  
Membrane Fraction ◽  
Microsomal Fraction ◽  
Membrane Vesicles ◽  
Cholesterol Content ◽  
Sucrose Density Gradient ◽  
Plasma Membrane Vesicles ◽  
Plasma Membrane Fraction

1. A plasma-membrane fraction was isolated from the smooth muscle of the pig stomach by using differential and sucrose-density-gradient centrifugations. When the centrifugation was carried out after preloading the crude microsomal fraction with Ca2+ in the presence of oxalate, the contamination of the plasma-membrane fraction by endoplasmic reticulum was decreased and a fraction enriched in endoplasmic reticulum vesicles filled with calcium oxalate crystals was obtained. 2. The plasmalemmal and endoplasmic-reticulum membranes could be distinguished by differences in the activity of marker enzymes and in the cholesterol content and by their different permeability to oxalate and phosphate. Oxalate and phosphate stimulated the Ca2+ uptake in the endoplasmic reticulum much more than in the plasmalemmal vesicles. In the plasma-membrane vesicles 40 mM-phosphate was more effective for stimulating the Ca2+ uptake than was 5 mM-oxalate, but the reverse was seen in the endoplasmic reticulum. 3. The high cholesterol/phospholipid ratio of the crude microsomal fraction are of the majority of the vesicles present in the crude microsomal fraction are of plasmalemmal origin. 4. The Ca2+ pump of the plasmalemmal and endoplasmic-reticulum vesicles could be differentiated by their different sensitivities to calmodulin. However, the two Ca2+-transport ATPases did not differ by their sensitivity to vanadate nor by the energization of the Ca2+ transport by different nucleoside triphosphates.

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