新型具有生物学用途的红色发光氯敏型荧光蛋白。

Department of Biochemistry and Molecular Biology II, Faculty of Pharmacy, Unidad de Excelencia en Quimica Aplicada a Biomedicina y Medioambiente (UEQ), University of Granada, Cartuja Campus, 18071 Granada, Spain.

Department of Physical Chemistry, Faculty of Pharmacy, Unidad de Excelencia en Quimica Aplicada a Biomedicina y Medioambiente (UEQ), University of Granada, C. U. Cartuja, 18071 Granada, Spain.

ACS Sens. 2021 Jul 23;6(7):2563-2573. doi: 10.1021/acssensors.1c00094. Epub 2021 Jun 20.

A new chloride-sensitive red fluorescent protein derived from is described. We found that mBeRFP exhibited moderate sensitivity to chloride and, via site-directed mutagenesis (S94V and R205Y), we increased the chloride affinity by more than an order of magnitude ( = 106 ± 6 mM) at physiological pH. In addition, isomerization of the chromophore produces a dual emission band with different chloride sensitivities, which allowed us to develop a ratiometric methodology to measure intracellular chloride concentrations.

本文描述了一种新型的氯离子敏感型红色荧光蛋白，来源于。我们发现 mBeRFP 对氯离子具有中等的敏感性，通过定点突变（S94V 和 R205Y），在生理 pH 值下，我们将氯离子的亲和力提高了一个数量级以上（= 106 ± 6 mM）。此外，生色团的异构化产生了一个具有不同氯离子敏感性的双发射带，这使我们能够开发出一种比率法来测量细胞内氯离子浓度。