Bioconjugation strategy for cell surface labelling with gold nanostructures designed for highly localized pH measurement.,Nature Communications

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Bioconjugation strategy for cell surface labelling with gold nanostructures designed for highly localized pH measurement.
Nature Communications ( IF 14.7 ) Pub Date : 2018-12-11 , DOI: 10.1038/s41467-018-07726-5
Leonardo Puppulin ₁ , Shigekuni Hosogi _{1,

2} , Hongxin Sun ₃ , Kazuhiko Matsuo ₄ , Toshio Inui _{5,

6} , Yasuaki Kumamoto ₇ , Toshinobu Suzaki ₈ , Hideo Tanaka ₇ , Yoshinori Marunaka _{1,

5,

9}

Affiliation

Department of Molecular Cell Physiology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kajii-cho, Kawaramachi-Hirokoji, Kyoto, 602-8566, Japan.
Department of Clinical and Translational Physiology, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto, 607-8414, Japan.
College of Pharmaceutical Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga, 525-8577, Japan.
Department of Anatomy and Developmental Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kajii-cho, Kawaramachi-Hirokoji, Kyoto, 602-8566, Japan.
Research Center for Drug Discovery and Pharmaceutical Development Science, Research Organization of Science and Technology, Ritsumeikan University, Kusatsu, 525-8577, Japan.
Saisei Mirai Clinics, Moriguchi, 3-34-8 Okubocho, Moriguchi-shi, Osaka, 570-0012, Japan.
Department of Pathology and Cell Regulation, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kajii-cho, Kawaramachi-Hirokoji, Kyoto, 602-8566, Japan.
Department of Biology, Graduate School of Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, 657-8501, Japan.
Research Institute for Clinical Physiology, Kyoto Industrial Health Association, 67 Kitatsuboi-cho, Nishino-kyo, Nakagyo-ku, Kyoto, 604-8472, Japan.

Regulation of intracellular pH is critically important for many cellular functions. The quantification of proton extrusion in different types of cells and physiological conditions is pivotal to fully elucidate the mechanisms of pH homeostasis. Here we show the use of gold nanoparticles (AuNP) to create a high spatial resolution sensor for measuring extracellular pH in proximity of the cell membrane. We test the sensor on HepG2 liver cancer cells and MKN28 gastric cancer cells before and after inhibition of Na⁺/H⁺ exchanger. The gold surface conjugation strategy is conceived with a twofold purpose: i) to anchor the AuNP to the membrane proteins and ii) to quantify the local pH from AuNP using surface enhanced Raman spectroscopy (SERS). The nanometer size of the cell membrane anchored sensor and the use of SERS enable us to visualize highly localized variation of pH induced by H⁺ extrusion, which is particularly upregulated in cancer cells.

中文翻译：

用于高度局部化 pH 测量的金纳米结构标记细胞表面的生物共轭策略。

细胞内 pH 值的调节对于许多细胞功能至关重要。不同类型细胞和生理条件下质子挤出的量化对于充分阐明 pH 稳态机制至关重要。在这里，我们展示了使用金纳米粒子 (AuNP) 来创建高空间分辨率传感器，用于测量细胞膜附近的细胞外 pH 值。我们在 Na ⁺ /H ⁺交换器抑制前后对 HepG2 肝癌细胞和 MKN28 胃癌细胞进行传感器测试。金表面缀合策略具有双重目的：i) 将 AuNP 锚定到膜蛋白上，ii) 使用表面增强拉曼光谱 (SERS) 量化 AuNP 的局部 pH 值。细胞膜锚定传感器的纳米尺寸和 SERS 的使用使我们能够可视化由 H ⁺挤出引起的高度局部的 pH 变化，这种变化在癌细胞中尤其上调。

更新日期：2018-12-11

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