Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior s/n, Ciudad Universitaria, 04510, México City, Mexico.
Departamento de Neurodesarrollo y Fisiología, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior S/n, Ciudad Universitaria, 04510, México City, Mexico.
Arch Biochem Biophys. 2024 Oct;760:110131. doi: 10.1016/j.abb.2024.110131. Epub 2024 Aug 21.
The effects of different pH incubation values and K on yeast plasma membrane potential (PMP) were studied both by the fluorescence changes and the accumulation of thioflavin T (ThT), a method that has been shown most adequate for both procedures. By the changes in fluorescence of ThT, the qualitative observation of PMP at the 3 evaluated pHs indicated that cells at pH 4.0 maintain a PMP lower, but close to the observed at pH 6.0 and 7.0. By measuring the accumulation of ThT and applying the Nernst equation on the different concentrations in and out, the values of PMP could also be estimated at the different pHs, resulting in values in mV, in agreement with our observations by following the fluorescence. Yeast cells at their native niches, or during fermentations must cope with low pHs, so the importance to maintain a robust PMP to survive. The contribution of bicarbonate, derived from the fermentation to the establishment of the PMP is also described. The experiments showed once more the efficacy of the methods used with this dye.
研究了不同 pH 值孵育值和 K 对酵母质膜电位(PMP)的影响,荧光变化和硫代黄素 T(ThT)的积累都证明了这两种方法是最适用的。通过 ThT 的荧光变化,对 3 个评估 pH 值下 PMP 的定性观察表明,pH 值为 4.0 的细胞维持的 PMP 较低,但接近 pH 值为 6.0 和 7.0 时观察到的 PMP。通过测量 ThT 的积累,并在内外不同浓度下应用能斯特方程,也可以估计不同 pH 值下的 PMP 值,以 mV 为单位,与我们通过荧光观察到的值一致。酵母细胞在其天然栖息地或发酵过程中必须应对低 pH 值,因此保持强大的 PMP 以维持生存至关重要。还描述了来自发酵的碳酸氢盐对 PMP 建立的贡献。实验再次证明了该染料所用方法的有效性。