Molecular characterization of a novel cell surface ADP-ribosyl cyclase from the sea urchin.
Molecular characterization of a novel cell surface ADP-ribosyl cyclase from the sea urchin.
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DOI：
10.1016/j.cellsig.2008.09.005
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发表时间：
2008-12
影响因子：
4.8
通讯作者：
Patel, Sandip
中科院分区：
文献类型：
--
作者：
Churaman, Dev;Boulware, Michael J.;Ramakrishnan, Latha;Geach, Timothy J.;Martin, Andrew C. R.;Vacquier, Victor D.;Marchant, Jonathan S.;Dale, Leslie;Patel, Sandip
海胆是一种广泛使用的模型系统，用于研究信使分子NAADP和环状ADP-核糖的钙信号传导。两者都是由ADP-核糖环化酶合成的，但我们对海胆中这些酶的分子理解是有限的。我们最近报道了海胆ADP-核糖基环化酶的一个大家族的克隆，并表明这些酶之一（SpARC 1）是活跃的内质网内腔内。这些研究表明，信使的产生是划分的。在这里，我们表征的SpARC 2的属性。SpARC 2催化NAADP和环状ADP-核糖的产生。不寻常的是，NAD替代物NGD是较差的底物。与SpARC 1相反，异源表达的SpARC 2通过糖基磷脂酰肌醇（GPI）-锚定位于质膜。SpARC 2的转录本在海胆卵中很容易检测到，并且发现大多数内源性膜结合活性是GPI锚定的。我们的数据揭示了海胆ADP-核糖基环化酶的性质的显着差异，并提供了进一步的证据，信使生产可能发生在胞质溶胶之外。
The sea urchin is an extensively used model system for the study of calcium signalling by the messenger molecules NAADP and cyclic ADP-ribose. Both are synthesized by ADP-ribosyl cyclases but our molecular understanding of these enzymes in the sea urchin is limited. We have recently reported the cloning of an extended family of sea urchin ADP-ribosyl cyclases and shown that one of these enzymes (SpARC1) is active within the endoplasmic reticulum lumen. These studies suggest that production of messengers is compartmentalized. Here we characterize the properties of SpARC2. SpARC2 catalysed both NAADP and cyclic ADP-ribose production. Unusually, the NAD surrogate, NGD was a poor substrate. In contrast to SpARC1, heterologously expressed SpARC2 localized to the plasma membrane via a glycosylphosphatidylinositol (GPI)-anchor. Transcripts for SpARC2 were readily detectable in sea urchin eggs and a majority of the endogenous membrane bound activity was found to be GPI-anchored. Our data reveal striking differences in the properties sea urchin ADP-ribosyl cyclases and provide further evidence that messenger production may occur outside of the cytosol.
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