Fernandes A R, Peixoto F P, Sá-Correia I
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1096 Lisboa Codex, Portugal.
Arch Microbiol. 1998 Dec;171(1):6-12. doi: 10.1007/s002030050671.
Cells of Saccharomyces cerevisiae exhibited a more active plasma membrane H+-ATPase during growth in media supplemented with CuSO4 concentrations equal to or below 1 mM than did cells cultivated in the absence of copper stress. Maximal specific activities were found with 0.5 mM CuSO4. ATPase activity declined when cells were grown with higher concentrations up to 1.5 mM (the maximal concentration that allowed growth), probably due to severe disorganization of plasma membrane. Cu2+-induced maximal activation was reflected in an increase of Vmax (approximately threefold) and in the slight decrease of the Km for MgATP (from 0.93 +/- 0.13 to 0.65 +/- 0.16 mM). The expression of the gene encoding the essential plasma membrane ATPase (PMA1) was reduced with a dose-dependent pattern in cells grown with Inhibitory concentrations of copper, while the weakly expressed PMA2 gene promoter was moderately more efficient in cells cultivated under mild copper stress (1.5-fold maximal activation). ATPase was activated by copper despite the slightly lower content of ATPase protein in the plasma membrane of Cu2+-grown cells and the powerful inhibitory effect of Cu2+ in vitro.
与在无铜胁迫条件下培养的细胞相比,酿酒酵母细胞在添加了浓度等于或低于1 mM的CuSO₄的培养基中生长时,其质膜H⁺-ATP酶表现出更活跃的状态。在0.5 mM CuSO₄时发现最大比活性。当细胞在高达1.5 mM(允许生长的最大浓度)的较高浓度下生长时,ATP酶活性下降,这可能是由于质膜严重紊乱所致。Cu²⁺诱导的最大激活反映在Vmax的增加(约三倍)以及MgATP的Km略有降低(从0.93±0.13降至0.65±0.16 mM)。在含有抑制浓度铜的细胞中,编码必需质膜ATP酶(PMA1)的基因表达呈剂量依赖性降低,而弱表达的PMA2基因启动子在轻度铜胁迫(最大激活1.5倍)下培养的细胞中效率适度更高。尽管在Cu²⁺生长的细胞的质膜中ATP酶蛋白含量略低,且Cu²⁺在体外具有强大的抑制作用,但ATP酶仍被铜激活。