Synthesis and biological testing of cell-permeant NAADP

细胞渗透性NAADP的合成及生物学测试

• 批准号: BB/D012694/1
• 负责人: Grant Churchill
• 金额: $ 27.14万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD012694%2F1
• 关键词: Synthesis; biological; testing; cell; permeant

Cells communicate with other cells and internally via chemical messengers. These messenger are molecules and exhibit features that make them uniquely recognizable to specialised receptors, or biological message centres. Messengers are made inside the cell membrane in response to outside stimuli, but cannot cross to or from the outside. This is because cells are enveloped by a membrane which keeps the contents in and potential contaminants outside. We are interested in a messenger molecule that increases calcium levels inside cells, called NAADP. Changes in calcium levels is what drives your heart to contract, your arm muscle to contract to raise a cup to your mouth and even the cells in your brain to talk to each other. Nobody is yet sure NAADP is working, so we want to get it into cells and find out. This messenger molecule does not naturally cross the cell membrane, so current state of the art techniques involve destroying the membrane then rebuilding it. This is difficult and requires specialised skills. We propose to try a different, more subtle approach by masking the molecules appearance, so that it can slip through the membrane. Once inside, the molecule sheds its mask, regains its appearance and provides its message. Getting NAADP across the cell membrane by stealth will provide us with new information about how this messenger works in different cells. Successful development of this new method will allow many more scientists to study this fascinating molecule.

细胞与其他细胞之间以及内部通过化学信使进行交流。这些信使是分子，它们的特征使它们能够被专门的受体或生物信息中心识别。信使是在细胞膜内对外界刺激做出反应，但不能与外界交叉。这是因为细胞被一层膜包裹着，把里面的东西和潜在的污染物都挡在了外面。我们感兴趣的是一种信使分子，它可以增加细胞内的钙水平，称为NAADP。钙水平的变化促使你的心脏收缩，你的手臂肌肉收缩，把杯子举到嘴里，甚至是你大脑中相互交谈的细胞。目前还没有人确定NAADP是否有效，所以我们希望将其移植到细胞中，并找出答案。这种信使分子不会自然地穿过细胞膜，所以目前最先进的技术包括破坏细胞膜，然后重建它。这是困难的，需要专门的技能。我们建议尝试一种不同的、更微妙的方法，通过掩盖分子的外观，使其能够滑过膜。一旦进入，分子就会脱下面具，重新露面，并提供信息。让NAADP秘密地穿过细胞膜，将为我们提供关于这种信使如何在不同细胞中发挥作用的新信息。这种新方法的成功开发将使更多的科学家能够研究这种迷人的分子。

项目成果

Synthesis of NAADP-AM as a Membrane-Permeant NAADP Analog

合成 NAADP-AM 作为膜渗透性 NAADP 类似物

• DOI: 10.1101/pdb.prot076927
• 发表时间: 2014
• 期刊: Cold Spring Harbor Protocols
• 作者: Galione A

Synthesis of caged NAADP.

笼中 NAADP 的合成。

• DOI: 10.1101/pdb.prot076943
• 发表时间: 2014
• 期刊: Cold Spring Harbor protocols
• 作者: Galione A

NAADP-mediated channel 'chatter' in neurons of the rat medulla oblongata

• DOI: 10.1042/bj20081138
• 发表时间: 2009-04-01
• 期刊: BIOCHEMICAL JOURNAL
• 影响因子: 4.1
• 作者: Brailoiu, G. Cristina;Brailoiu, Eugen;Dun, Nae J.
• 通讯作者: Dun, Nae J.

Synthesis of [³²P]NAADP for the radioreceptor binding assay.

用于放射受体结合测定的[32P]NAADP 的合成。

• DOI: 10.1101/pdb.prot076919
• 发表时间: 2014
• 期刊: Cold Spring Harbor protocols
• 作者: Galione A