Organic Chemistry, University of Hamburg, 20146, Hamburg, Germany.
The Calcium Signalling Group, Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, 20246, Hamburg, Germany.
Nat Commun. 2024 Sep 13;15(1):8008. doi: 10.1038/s41467-024-52024-y.
Upon stimulation of membrane receptors, nicotinic acid adenine dinucleotide phosphate (NAADP) is formed as second messenger within seconds and evokes Ca signaling in many different cell types. Here, to directly stimulate NAADP signaling, MASTER-NAADP, a Membrane permeAble, STabilized, bio-rEversibly pRotected precursor of NAADP is synthesized and release of its active NAADP mimetic, benzoic acid C-nucleoside, 2'-phospho-3'F-adenosine-diphosphate, by esterase digestion is confirmed. In the presence of NAADP receptor HN1L/JPT2 (hematological and neurological expressed 1-like protein, HN1L, also known as Jupiter microtubule-associated homolog 2, JPT2), this active NAADP mimetic releases Ca and increases the open probability of type 1 ryanodine receptor. When added to intact cells, MASTER-NAADP initially evokes single local Ca signals of low amplitude. Subsequently, also global Ca signaling is observed in T cells, natural killer cells, and Neuro2A cells. In contrast, control compound MASTER-NADP does not stimulate Ca signaling. Likewise, in cells devoid of HN1L/JPT2, MASTER-NAADP does not affect Ca signaling, confirming that the product released from MASTER-NAADP is a bona fide NAADP mimetic.
当细胞膜受体受到刺激时，烟酰胺腺嘌呤二核苷酸磷酸（NAADP）在几秒钟内形成第二信使，并在许多不同的细胞类型中引发 Ca 信号。在这里，为了直接刺激 NAADP 信号，合成了 MASTER-NAADP，一种透膜、稳定、生物可逆保护的 NAADP 前体，通过酯酶消化证实其活性 NAADP 类似物苯甲酸 C-核苷 2'-磷酸-3'F-腺苷二磷酸的释放。在存在 NAADP 受体 HN1L/JPT2（血液和神经表达 1 样蛋白，HN1L，也称为木星微管相关同源物 2，JPT2）的情况下，这种活性 NAADP 类似物释放 Ca 并增加 1 型肌醇 1,4,5-三磷酸受体的开放概率。当添加到完整细胞时，MASTER-NAADP 最初会引发低幅度的单个局部 Ca 信号。随后，在 T 细胞、自然杀伤细胞和 Neuro2A 细胞中也观察到全局 Ca 信号。相比之下，对照化合物 MASTER-NADP 不会刺激 Ca 信号。同样，在没有 HN1L/JPT2 的细胞中，MASTER-NAADP 不会影响 Ca 信号，这证实从 MASTER-NAADP 释放的产物是真正的 NAADP 类似物。
