Okuyama Akari, Hososhima Shoko, Kandori Hideki, Tsunoda Satoshi P
Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Nagoya, Aichi, Japan.
Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Nagoya, Aichi, Japan; OptoBioTechnology Research Center, Nagoya Institute of Technology, Nagoya, Aichi, Japan.
Biophys J. 2024 Dec 17;123(24):4274-4284. doi: 10.1016/j.bpj.2024.09.007. Epub 2024 Sep 6.
Proton-pumping rhodopsins are light-driven proton transporters that have been discovered from various microbiota. They are categorized into two groups: outward-directed and inward-directed proton pumps. Although the directions of transport are opposite, they are active proton transporters that create an H gradient across a membrane. Here, we aimed to study the driving force of the proton-pumping rhodopsins and the effect of ΔΨ and ΔpH on their pumping functions. We systematically characterized the H transport properties of nine different rhodopsins, six outward-directed H pumps and three inward-directed pumps, by patch-clamp measurements after expressing them in mammalian cells. The driving force of each pump was estimated from the slope of the current-voltage relations (I-V plot). Notably, among the tested rhodopsins, we found a large variation in driving forces, ranging from 83 to 399 mV. The driving force and decay rate of each pump current exhibited a good correlation. We determined driving forces under various pHs. pH dependency was less than predicted by the Nernst potential in most of the rhodopsins. Our study demonstrates that the H-pumping rhodopsins from different organisms exhibit various pumping properties in terms of driving force, kinetics, and pH dependency, which could be evolutionarily derived from adaptations to their environments.
质子泵视紫红质是从各种微生物群中发现的光驱动质子转运体。它们分为两类:外向型和内向型质子泵。尽管运输方向相反,但它们都是活跃的质子转运体,能在膜上形成H+梯度。在此,我们旨在研究质子泵视紫红质的驱动力以及膜电位差(ΔΨ)和pH梯度(ΔpH)对其泵浦功能的影响。我们通过在哺乳动物细胞中表达后采用膜片钳测量技术,系统地表征了九种不同视紫红质的H+转运特性,其中包括六种外向型H+泵和三种内向型泵。根据电流-电压关系曲线(I-V曲线)的斜率估算每种泵的驱动力。值得注意的是,在所测试的视紫红质中,我们发现驱动力存在很大差异,范围从83到399 mV。每种泵电流的驱动力和衰减速率呈现出良好的相关性。我们测定了在各种pH值下的驱动力。在大多数视紫红质中,pH依赖性小于能斯特电位预测的值。我们的研究表明,来自不同生物体的H+泵视紫红质在驱动力、动力学和pH依赖性方面表现出各种泵浦特性,这些特性可能是在进化过程中适应其环境而形成 的。