Missiaen L, Wuytack F, De Smedt H, Vrolix M, Casteels R
Department of Physiology, Catholic University of Leuven, Belgium.
Biochem J. 1988 Aug 1;253(3):827-33. doi: 10.1042/bj2530827.
The only known cellular action of AlF4- is to stimulate the G-proteins. The aim of the present work is to demonstrate that AlF4- also inhibits 'P'-type cation-transport ATPases. NaF plus AlCl3 completely and reversibly inhibits the activity of the purified (Na+ + K+)-ATPase (Na+- and K+-activated ATPase) and of the purified plasmalemmal (Ca2+ + Mg2+)-ATPase (Ca2+-stimulated and Mg2+-dependent ATPase). It partially inhibits the activity of the sarcoplasmic-reticulum (Ca2+ + Mg2+)-ATPase, whereas it does not affect the mitochondrial H+-transporting ATPase. The inhibitory substances are neither F- nor Al3+ but rather fluoroaluminate complexes. Because AlF4- still inhibits the ATPase in the presence of guanosine 5'-[beta-thio]diphosphate, and because guanosine 5'-[beta gamma-imido]triphosphate does not inhibit the ATPase, it is unlikely that the inhibition could be due to the activation of an unknown G-protein. The time course of inhibition and the concentrations of NaF and AlCl3 required for this inhibition differ for the different ATPases. AlF4- inhibits the (Na+ + K+)-ATPase and the plasmalemmal (Ca2+ + Mg2+)-ATPase noncompetitively with respect to ATP and to their respective cationic substrates, Na+ and Ca2+. AlF4- probably binds to the phosphate-binding site of the ATPase, as the Ki for inhibition of the (Na+ + K+)-ATPase and of the plasmalemmal (Ca2+ + Mg2+)-ATPase is shifted in the presence of respectively 5 and 50 mM-Pi to higher concentrations of NaF. Moreover, AlF4- inhibits the K+-activated p-nitrophenylphosphatase of the (Na+ + K+)-ATPase competitively with respect to p-nitrophenyl phosphate. This AlF4- -induced inhibition of 'P'-type cation-transport ATPases warns us against explaining all the effects of AlF4- on intact cells by an activation of G-proteins.
已知AlF4-唯一的细胞作用是刺激G蛋白。本研究的目的是证明AlF4-也抑制“P”型阳离子转运ATP酶。NaF加AlCl3能完全且可逆地抑制纯化的(Na+ + K+)-ATP酶(Na+和K+激活的ATP酶)以及纯化的质膜(Ca2+ + Mg2+)-ATP酶(Ca2+刺激且Mg2+依赖的ATP酶)的活性。它部分抑制肌浆网(Ca2+ + Mg2+)-ATP酶的活性,而不影响线粒体H+转运ATP酶。抑制物质既不是F-也不是Al3+,而是氟铝酸盐复合物。由于在存在鸟苷5'-[β-硫代]二磷酸的情况下AlF4-仍能抑制ATP酶,并且由于鸟苷5'-[βγ-亚氨基]三磷酸不抑制ATP酶,所以这种抑制不太可能是由于激活未知的G蛋白所致。不同ATP酶的抑制时间进程以及产生这种抑制所需的NaF和AlCl3浓度各不相同。AlF4-对(Na+ + K+)-ATP酶和质膜(Ca2+ + Mg2+)-ATP酶的抑制作用相对于ATP及其各自的阳离子底物Na+和Ca2+而言是非竞争性的。AlF4-可能与ATP酶的磷酸结合位点结合,因为在分别存在5 mM和50 mM无机磷酸的情况下,抑制(Na+ + K+)-ATP酶和质膜(Ca2+ + Mg2+)-ATP酶的抑制常数Ki会向更高浓度的NaF偏移。此外,AlF4-相对于对硝基苯磷酸竞争性抑制(Na+ + K+)-ATP酶的K+激活的对硝基苯磷酸酶。AlF4-诱导的对“P”型阳离子转运ATP酶的这种抑制作用提醒我们,不能仅通过G蛋白的激活来解释AlF4-对完整细胞的所有作用。