Novel microsomal anion-sensitive Mg2+-ATPase activity in rat brain

Inagaki, Chiyoko; Tanaka, Toshizo; Hara, Mitsuyoshi; Ishiko, Jiro

doi:10.1016/0006-2952(85)90638-0

Cited by 51 publications

(21 citation statements)

References 21 publications

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“…The superiority of the detergent combination has been reported in the solubilization of ATPases from renal brush-border membranes (16) and thermophilic cyanobacteria (17). As previously described (10,12), Mg2+ ATPases in the microsomes were inhibited by EA biphasically with a deflecting point at 0.3-0.4 mM. In the present study, the solubilized Mgt+-ATPases exhibited these properties with respect to the sensitivity to EA.…”

Section: Discussionsupporting

confidence: 82%

“…We previously showed that brain micro somes contain two types of Mgt+-ATPases activities: ethacrynic acid (EA) highly sensitive and EA less sensitive Mgt+-ATPase activities (10)(11)(12)(13).…”

mentioning

confidence: 99%

“…The former activity was mainly located in plasma membrane fractions and was sensitive to anions, especially to Cl (a stimulatory anion) and NO3 (an inhibitory anion), and the latter was insensitive to the anions and rather widely distributed (12,13 In the combined detergent-treatment, CHAPS was added to the suspension of microsomes (10 mg protein/ml) to achieve a final CHAPS concentration of 10 mM, and the mixture was centrifuged at 20,000 x g for 15 min. To the resulting supernatant, OcGlu up to 50 mM was added.…”

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confidence: 99%

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Solubilization and separation of ethacrynic acid(EA) highly sensitive and EA less sensitive Mg2+-ATPases in the rat brain.

et al. 1987

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Abstract-Rat brain microsomal Mgt+-ATPases with two distinct activities: ethacrynic acid (EA) highly sensitive and EA less sensitive Mgt+-ATPase activities were solubilized by the combined treatment with 10 mM 3-(3-chlolamidopropyl) dimethylammonio-1 -propane-sulfate (CHAPS) and 30 mM octyl-Q-D-glucoside. The solubilized enzymes had properties similar to those of the membrane-bound enzyme in microsomes with respect to the sensitivity to EA and Cl-, although the optimal pH and the affinity to ATP were slightly altered after the solubilization. Fast protein liquid chromatography of the solubilized enzymes on an anion exchanger (Mono Q) column with a linear NaCI gradient (0-1.0 M) yielded separate peaks for EA highly sensitive and EA less sensitive Mgt+-ATPase activities at 0.1 and 0.35 M NaCI, respectively.Polyacrylamide gradient gel electrophoresis of the samples from the peak-fractions of EA highly sensitive and EA less sensitive Mgt+-ATPase activities yielded prominent bands at 600 and 70 kDa, respectively. These results indicate that EA highly sensitive Mgt+-ATPase is solubilized and separated from EA less sensitive Mgt+-ATPase as a large enzyme molecule with anion-sensitive sites.

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Section: Discussionsupporting

confidence: 82%

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confidence: 99%

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confidence: 99%

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Solubilization and separation of ethacrynic acid(EA) highly sensitive and EA less sensitive Mg2+-ATPases in the rat brain.

et al. 1987

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“…Another mechanism to help maintain the Cl Ϫ transmembrane gradient is the Cl Ϫ -ATPase, a plasma membrane ion pump that extrudes Cl Ϫ from neurons (Inagaki et al, 1985;Shiroya et al, 1989). Because there is incomplete recovery of ATP on reoxygenation (Galeffi et al, 2000), we determined whether the accumulation of intracellular Cl Ϫ after OGD was attributable to a reduction in Cl Ϫ -ATPase activity.…”

Section: Effect Of Ogd On CL ؊ -Atpase Activitymentioning

confidence: 99%

“…In addition, inhibition of NKCC1 is neuroprotective after anoxia or OGD (Müller, 2000;Pond et al, 2004) and in rats after focal ischemia (Yan et al, 2001). In the case of Cl Ϫ extrusion mechanisms, the KCC2 and the Cl Ϫ -ATPase help to maintain the Cl Ϫ gradient in adult neurons (Inagaki et al, 1985;Shiroya et al, 1989;Rivera et al, 1999); they extrude Cl Ϫ in response to GABA A receptor-activated Cl Ϫ channel opening. KCC2 is regulated in a variety of ways after different types of brain insults.…”

Section: Ogd Effects On Intracellular CL ؊ and Synaptic Transmissionmentioning

confidence: 99%