• Title/Summary/Keyword: $Ca^{++}$uptake

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Characterization of Microsomal $Ca^{2+}$ Uptake in Tomato Root Tissues (토마토 뿌리조직에서 분리한 마이크로솜의 $Ca^{2+}$ 흡수 특성)

  • Cho, Kwang-Hyun;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.116-122
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    • 1999
  • In order to characterize the property of $Ca^{2+}$ transport in plant cell, microsomes were prepared from the roots of tomato and microsomal $^{45}Ca^{2+}$ uptake was measured. When 1 mM vanadate, a selective inhibitor of P-type ATPases, 50 mM $NO_3^-$, a specific inhibitor of vacuolar $H^{+}-ATPase$, and both of these inhibitors were treated, the microsomal $^{45}Ca^{2+}$ uptakes were inhibited by 20, 33 and 47%, respectively. The inhibitory effects of these two inhibitors were investigated by using a protonophore, gramicidin. When the chemical gradient of $H^{+}$ was relieved by gramicidin, the uptake was decreased by 30%, implying the presence of $Ca^{2+}/H^+$ antiporter in the microsomal membrane. In the $^{45}Ca^{2+}$ uptake experiment, the effect of gramicidin was independent of vanadate-induced inhibition. However, when the activity of vacuolar $H^{+}-ATPase$ was inhibited by $NO_3^-$, the effect of gramicidin was severely decreased. Meanwhile, thapsigargin, a specific antagonist of ER/SR-type $Ca^{2+}-ATPase$, inhibited the microsomal $^{45}Ca^{2+}$ uptake and the maximum inhibitory effect was obtained at $10\;{\mu}M$. The effect of thapsigargin was blocked by $NO_3^-$ and gramicidin, but not by vanadate. These results imply that vanadate directly inhibits the activity of $Ca^{2+}-ATPase$; however, $NO_3^-$ and thapsigargin block the activity of $Ca^{2+}/H^+$ antiporter by inhibiting the vacuolar $H^{+}-ATPase$. In conclusion, the microsomal $^{45}Ca^{2+}$ uptakes are mediated by two major enzymes, $Ca^{2+}-ATPase$ and $Ca^{2+}/H^+$ antiporter in tomato root tissue.

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Effect of a Phospholamban Peptide on the Skeletal Sarcoplasmic Reticulum $Ca^{2+}$ Transport (골격근 근장그물 칼슘이동에 대한 Phospholamban 펩타이드의 조절)

  • Kim, Hae-Won;Lee, Hee-Ran
    • The Korean Journal of Pharmacology
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    • v.30 no.1
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    • pp.117-124
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    • 1994
  • Phospholamban is the regulator of $Ca^{2+}-ATPase$ in cardiac sarcoplasmic reticulum(SR). The mechanism of regulation appears to involve inhibition by dephosphorylated phospholamban. Phosphorylation of phospholamban relieves this inhibition. Recently, there has been a report that the cytoplasmic domain (amino acids 1-25) of phospholamban is insufficient to inhibit the $Ca^{2+}$ pump. To explore the domains of phospholamban responsible for $Ca^{2+}-ATPase$ inhibitory activity, we examined the effect of a synthetic phospholamban peptide consisting of amino acid residues 1-25 on $Ca^{2+}$ uptake by reconstituted skeletal SR $Ca^{2+}-ATPase$. The $Ca^{2+}-ATPase$ of skeletal SR was purified and reconstituted in proteoliposomes containing phosphatidylcholine (PC) or phosphatidylcholine: phosphatidylserine (PC:PS). Inclusion of a phospholamban peptide in PC proteoliposomes was associated with significant inhibition of the initial rates of $Ca^{2+}$ uptake at pCa 6.0, and phosphorylation of this peptide by the catalytic subunit of cAMP-dependent protein kinase reversed the inhibitory effect on the $Ca^{2+}$ pump. Similar effects of phospholamban peptide were also observed using PC:PS proteoliposomes. Based on these results, we could conclude that the cytoplasmic domain of phospholamban, containing the phosphorylation sites, by itself is sufficient to inhibit the $Ca^{2+}$ pump of SR.

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Mitochondrial Ca2+ Uptake Relieves Palmitate-Induced Cytosolic Ca2+ Overload in MIN6 Cells

  • Ly, Luong Dai;Ly, Dat Da;Nguyen, Nhung Thi;Kim, Ji-Hee;Yoo, Heesuk;Chung, Jongkyeong;Lee, Myung-Shik;Cha, Seung-Kuy;Park, Kyu-Sang
    • Molecules and Cells
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    • v.43 no.1
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    • pp.66-75
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    • 2020
  • Saturated fatty acids contribute to β-cell dysfunction in the onset of type 2 diabetes mellitus. Cellular responses to lipotoxicity include oxidative stress, endoplasmic reticulum (ER) stress, and blockage of autophagy. Palmitate induces ER Ca2+ depletion followed by notable store-operated Ca2+ entry. Subsequent elevation of cytosolic Ca2+ can activate undesirable signaling pathways culminating in cell death. Mitochondrial Ca2+ uniporter (MCU) is the major route for Ca2+ uptake into the matrix and couples metabolism with insulin secretion. However, it has been unclear whether mitochondrial Ca2+ uptake plays a protective role or contributes to lipotoxicity. Here, we observed palmitate upregulated MCU protein expression in a mouse clonal β-cell, MIN6, under normal glucose, but not high glucose medium. Palmitate elevated baseline cytosolic Ca2+ concentration ([Ca2+]i) and reduced depolarization-triggered Ca2+ influx likely due to the inactivation of voltage-gated Ca2+ channels (VGCCs). Targeted reduction of MCU expression using RNA interference abolished mitochondrial superoxide production but exacerbated palmitate-induced [Ca2+]i overload. Consequently, MCU knockdown aggravated blockage of autophagic degradation. In contrast, co-treatment with verapamil, a VGCC inhibitor, prevented palmitate-induced basal [Ca2+]i elevation and defective [Ca2+]i transients. Extracellular Ca2+ chelation as well as VGCC inhibitors effectively rescued autophagy defects and cytotoxicity. These observations suggest enhanced mitochondrial Ca2+ uptake via MCU upregulation is a mechanism by which pancreatic β-cells are able to alleviate cytosolic Ca2+ overload and its detrimental consequences.

Charateristics of Voltage Dependent Calcium Uptake and Norepinephrine Release in Hypothalamus of DOCA-salt Hypertensive Rats

  • Lee, Jean-Young;Kim, Hae-Jung;Jung, Eun-Young;Chung, Hye-Joo;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • v.1 no.2
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    • pp.171-176
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    • 1993
  • Purpose of the present study was to clarify the role of noradrenergic neural activities in hypothalamus for either triggering or maintaining hypertension in deoxycorticosterone (DOCA)-salt hypertensive rats. Two groups of animals were prepared: 1) normotensive Wistar rats and 2) DOCA-salt induced hypertensive rats. Voltage dependent $^{45}Ca^{++}$ uptake, endogenous norepinephrine release, and the catecholamine content in the hypothalamus of DOCA-salt hypertensive and normotensive Wistar rats were compared. Animals at 4, 6 and 16 week-old of two groups were sacrificed by decapitation and hypothalamus was dissected out. Voltage dependent calcium uptake and norepinephrine release were determined from hypothalamic synaptosomes either in low potassium or high potassium stimulatory condition by using $^{45}Ca^{++}$ isotope and HPLC-ECD technique. Degrees of voltage dependent $^{45}Ca^{++}$ uptake and norepinephrine release in hypothalamic synaptosomes of 16-week-old DOCA-salt hypertensive rats were significantly greater than those of age matched normotensive control rats. The norepinephrine and dopamine contents of hypothalamus were about the same in two groups of animals. These results suggest that the alteration of evoked norepinephrine release related to calcium uptake in hypothalamus may play a role in the maintenance of hypertension in DOCA-salt hypertensive rats.

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Evaluation on the Phosphate Uptake Rate of Green Algae under Diurnal Rhythm and Algae Control Method (일주기에 따른 녹조류의 인흡수율 및 조류제어방법 평가)

  • Jang Kam-Yong;Lim Kyoung-Mook;Noh Tae-Mok;Lee Hac-Su;Park Sung-Ha
    • Journal of Environmental Science International
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    • v.14 no.9
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    • pp.873-879
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    • 2005
  • This study was investigate to evaluate the phosphate uptake rate of green algae in relation to diurnal rhythm and algae control method. The phosphate uptake rates of Chlorella vulgaris and Ankistrodesmus convolutus increased in light period and decreased in dark period. On the contrary, those of Chlamydomonas sp. showed a peak in the late dark period. The differences among species in phosphate uptake in relation to diurnal rhythm were due to the severe competition among species and seemed to alleviate the competition for nutrient supplies. The compound of CellCaSi, Ca and Fe showed the effective removal of the phosphorus. The extracts from rice and barley straw exhibited a significant effect on the growth inhibition of Microcystis aeruginosa.

Effect of Propranolol on the $Ca^{++}$-regulation of Cardiac Sarcoplasmic Reticulum and Mitochondria (Propranolol이 심근 sarcoplasmic reticulum 및 mitochondria 의 $Ca^{++}$ 조절작용에 미치는 효과에 관한 연구)

  • 최수승
    • Journal of Chest Surgery
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    • v.19 no.2
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    • pp.197-208
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    • 1986
  • Propranolol is one of clinically useful antiarrhythmic agents and electrophysiologically classified as group II. And the negative inotropic effect which is not related to adrenolytic effect has been demonstrated with high concentration of propranolol. On the other hand, it has been well known that the calcium plays a central role in excitation-contraction coupling process of myocardium and also in electrophysiological changes of cell membrane. Author studies the effect of propranolol on calcium uptake and release in sarcoplasmic reticulum and mitochondria prepared from porcine myocardium to investigate the mechanism of action of propranolol on myocardium. The results are summarized as follow: 1] The maximum Ca++-uptake of sarcoplasmic reticulum is inhibited by propranolol in a dose dependent manner. 2] The release of calcium from sarcoplasmic reticulum is not affected by propranolol but with higher than 1x10-3 M of propranolol, rate of calcium release from sarcoplasmic reticulum is decreased. 3] Propranolol inhibits the maximum uptake and uptake rate of calcium in mitochondria non-competitively. [Ki = 6.21 x 10-4 M] 4] The rate of Na+ induced calcium release from mitochondrion shows a function of [Na+]2 and is inhibited by propranolol with the concentration significantly lower than that affect the calcium uptake in sarcoplasmic reticulum and in mitochondria [Ki = 2.91 x 10-5 M]. These results suggest that propranolol affects the intracellular calcium homeostasis which may considered to be one of the mechanism of action of propranolol on myocardium.

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Studies on the Transport of Organic Acids in the Rabbit Kindey Slice, with Special Reference to the Role of Various Electrolytes (가토신피질절편(家兎腎皮質切片)에서의 유기산이동(有機酸移動)에 관(關)한 연구(硏究) -특(特)히 전해질(電解質)의 영향(影響)에 대(對)하여-)

  • Chung, Soon-Tong
    • The Korean Journal of Physiology
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    • v.2 no.1
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    • pp.59-71
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    • 1968
  • The uptake of phenolsulfonphthalein (PSP) and of paraaminohippuric acid (PAH) by cortical slices of the rabbit kidney was investigated while varying the composition of medium. The overall uptake of these substances displayed typical active transport characteristics and was significantly enhanced in presence of acetate. When the phosphate buffer was used the optimal pH was 7.4 for both substances. However, when the tris-buffer was used the optimal pH was 7.4 for PSP and 8.3 for PAH. Removal of $Na^+$ from the medium resulted in a significant reduction in the uptake. Similar results, though lesser in magnitude, were obtained when either $K^+\;or\;Ca^{++}$ was removed from the medium. However, there was no additive effect when $K^+\;and/or\;Ca^{++}$ were additionally removed from the $Na^+-free$ medium. The presence of ${NH_4}^+$ greatly reduced while $Li^+\;and\;Mg^{++}$ moderately reduced the uptake of both substances. However, choline had no effect. In substrate-leached slices, acetate greatly enhance the uptake of organic acids; but this action was not demonstrable in absence of $Na^+,\;K^+\;or\;Ca^{++}$.

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Calcium Ion Effect on the Sugar-H+ -Cotransport System in Chlorella vulgaris (Chlorella vulgaris의 당류 능동수송계에 미치는 칼슘 이온의 영향)

  • 조봉희
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.321-326
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    • 1993
  • Sugar uptake is accompanied with H+-substrate-symport generally. Both H+/sugar-and H+/K+ stoichiometries during the sugar-uptake have been reported to be exactly 1 : 1. This paper reports that the stoichiometries were enhanced dramatically by the addition of CaCl2 into the medium and by the high cell density of 200 $\mu$L pc/mL. The concentration of free Ca2+ ions in the cells increased significantly with cell density. It is suggested that the free Ca2+ ions are responsible for the change of stoichiometry of sugar transport system by regulation of H+ ion level of biomembrane.

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Effects of Poisonous Mushroom Extracts on the Microsomal $^{45}Ca^{2+}$ Uptake and Release in Porcine Epithelial Cells

  • Cho, Kyoung-Soo;Hwang, Young-Soo;Kim, Young-Kee
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.39-39
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    • 1996
  • Extracts were prepared from six different poisonous mushrooms in order to identify biologically active natural ligands. The effects of these extracts were examined on the microsomal $^{45}$ Ca$^{2+}$ uptake and $^{45}$ Ca$^{2+}$ release. Five out of six species apparently inhibited the activity of total microsomal ATPases prepared from the epithelial cells of pig airway. (omitted)ted)

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Agricultural Methods for Toxicity Alleviation in Metal Contaminated Soils: A Review

  • Arunakumara, Kkiu;Walpola, Buddhi Charana;Yoon, Min-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.46 no.2
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    • pp.73-80
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    • 2013
  • Due to the fact that possible risk associated with soil-crop-food chain transfer, metal contamination in croplands has become a major topic of wide concern. Accumulation of toxic metals in edible parts of crops grown in contaminated soils has been reported from number of crops including rice, soybean, wheat, maize, and vegetables. Therefore, in order to ensure food safety, measures are needed to be taken in mitigating metal pollution and subsequent uptake by crop plants. Present paper critically reviewed some of the cost effective remediation techniques used in minimizing metal uptake by crops grown in contaminated soils. Liming with different materials such as limestone ($CaCO_3$), burnt lime (CaO), slaked lime [$Ca(OH)_2$], dolomite [$CaMg(CO_3)_2$], and slag ($CaSiO_3$) has been widely used because they could elevate soil pH rendering metals less-bioavailable for plant uptake. Zn fertilization, use of organic amendments, crop rotation and water management are among the other techniques successfully employed in reducing metal uptake by crop plants. However, irrespectively the mitigating measure used, heterogeneous accumulation of metals in different crop species is often reported. The inconsistency might be attributed to the genetic makeup of the crops for selective uptake, their morphological characteristics, position of edible parts on the plants in respect of their distance from roots, crop management practices, the season and to the soil characteristics. However, a sound conclusion in this regard can only be made when more scientific evidence is available on case-specific researches, in particular from long-term field trials which included risks and benefits analysis also for various remediation practices.