基于GSK-7975A的化学工具的开发,用于研究储存操作的钙进入细胞

IF 4.3 2区 生物学 Q2 CELL BIOLOGY
Dominic Tscherrig , Rajesh Bhardwaj , Daniel Biner , Jan Dernič , Daniela Ross-Kaschitza , Christine Peinelt , Matthias A. Hediger , Martin Lochner
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引用次数: 0

摘要

许多生理功能,如细胞分化、增殖、肌肉收缩、神经传递和受精,都受Ca2+水平变化的调节。细胞内主要的Ca2+储存是内质网(ER)。某些细胞过程诱导内质网储存耗尽,例如通过激活IP3受体,进而诱导储存再填充过程,称为储存操作钙进入(SOCE)。这种再填充过程涉及内质膜中Ca2+感应基质相互作用分子(STIM)和质膜中的Orai蛋白之间的蛋白-蛋白相互作用。完全组装的STIM/Orai复合物然后形成高度选择性的Ca2+通道,称为Ca2+释放激活的Ca2+通道(CRAC), Ca2+离子通过该通道流入细胞质,随后被肌浆/内质网钙atp酶(SERCA)泵入内质网。异常的SOCE与许多人类疾病和癌症有关,因此主要参与者STIM和Orai引起了重大的治疗兴趣。已经开发了几种有效的实验和临床候选化合物,并有助于研究各种细胞类型中的SOCE。我们基于已知的SOCE抑制剂GSK-7975A合成了多种新型小分子探针。在这里,我们提出GSK-7975A衍生物,具有光笼化,光交联,生物素和可点击的部分,也含有氘标签。对这些GSK-7975A探针使用荧光成像板读取器(FLIPR)-基于四氢钙离子成像分析的评估显示,大多数合成修饰对SOCE抑制活性没有有害影响。光笼式GSK-7975A也用于膜片钳电生理实验。总之,我们已经开发了许多活性的、基于gsk -7975的分子探针,这些探针具有有趣的特性,因此是在各种细胞和环境中研究SOCE的有用实验工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Development of chemical tools based on GSK-7975A to study store-operated calcium entry in cells

Development of chemical tools based on GSK-7975A to study store-operated calcium entry in cells

Many physiological functions, such as cell differentiation, proliferation, muscle contraction, neurotransmission and fertilisation, are regulated by changes of Ca2+ levels. The major Ca2+ store in cells is the endoplasmic reticulum (ER). Certain cellular processes induce ER store depletion, e.g. by activating IP3 receptors, that in turn induces a store refilling process known as store-operated calcium entry (SOCE). This refilling process entails protein-protein interactions between Ca2+ sensing stromal interaction molecules (STIM) in the ER membrane and Orai proteins in the plasma membrane. Fully assembled STIM/Orai complexes then form highly selective Ca2+ channels called Ca2+ release-activated Ca2+ Channels (CRAC) through which Ca2+ ions flow into the cytosol and subsequently are pumped into the ER by the sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA). Abnormal SOCE has been associated with numerous human diseases and cancers, and therefore key players STIM and Orai have attracted significant therapeutic interest. Several potent experimental and clinical candidate compounds have been developed and have helped to study SOCE in various cell types. We have synthesized multiple novel small-molecule probes based on the known SOCE inhibitor GSK-7975A. Here we present GSK-7975A derivatives, which feature photo-caging, photo-crosslinking, biotin and clickable moieties, and also contain deuterium labels. Evaluation of these GSK-7975A probes using a fluorometric imaging plate reader (FLIPR)-Tetra-based Ca2+ imaging assay showed that most synthetic modifications did not have a detrimental impact on the SOCE inhibitory activity. The photo-caged GSK-7975A was also used in patch-clamp electrophysiology experiments. In summary, we have developed a number of active, GSK-7975A-based molecular probes that have interesting properties and therefore are useful experimental tools to study SOCE in various cells and settings.

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来源期刊
Cell calcium
Cell calcium 生物-细胞生物学
CiteScore
8.70
自引率
5.00%
发文量
115
审稿时长
35 days
期刊介绍: Cell Calcium covers the field of calcium metabolism and signalling in living systems, from aspects including inorganic chemistry, physiology, molecular biology and pathology. Topic themes include: Roles of calcium in regulating cellular events such as apoptosis, necrosis and organelle remodelling Influence of calcium regulation in affecting health and disease outcomes
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