Imaging and manipulating phosphoinositides in living cells
Imaging and manipulating phosphoinositides in living cells
复制标题
DOI：
10.1113/jphysiol.2007.132795
复制
发表时间：
2007-08-01
影响因子：
5.5
通讯作者：
Balla, Tamas
中科院分区：
文献类型：
--
作者：
Balla, Tamas
磷脂肌醇是几乎所有生物膜的次要磷脂成分，但它们在控制膜结合信号传导事件中发挥着重要作用。磷酸肌醇是由磷脂酰肌醇 (PtdIns) 产生的，通过肌醇脂质激酶的不同家族将位于膜细胞质界面的肌醇首基的三个位置（3、4 和 5）中的一个或多个位置磷酸化。有趣的是，即使在简单的生物体中，许多激酶反应也由不止一种形式的激酶催化，并且这些酶通常具有非冗余功能。肌醇磷酸酶也存在类似的多样性，这种酶以一定程度的特异性使磷酸肌醇去磷酸化，其损伤通常与人类疾病有关。酶水平上的这种程度的多样性以及这些脂质在细胞调节中的普遍作用假设肌醇脂质在空间和功能上受到特定膜区室的限制。在细胞水平上研究这些脂质的作用是一个重大的方法学挑战。在过去的 10 年里，在创建可通过荧光或共聚焦显微镜监测活细胞中肌醇脂质变化的试剂方面取得了重大进展。还正在开发新的方法来以受控的方式操纵特定膜室中的这些脂质。本文回顾了肌醇研究的一些历史方面，并描述了新的方法论进展，强调了它们的巨大潜力以及使用它们时可能遇到的问题。
Phosphoinositides are minor phospholipid constituents of virtually every biological membrane yet they play fundamental roles in controlling membrane-bound signalling events. Phosphoinositides are produced from phosphatidylinositol (PtdIns) by phosphorylation of one or more of three positions (3, 4 and 5) of the inositol headgroup located at the membrane cytoplasmic interface by distinct families of inositol lipid kinases. Intriguingly, many of the kinase reactions are catalysed by more than one form of the kinases even in simple organisms and these enzymes often assume non-redundant functions. A similar diversity is seen with inositide phosphatases, the enzymes that dephosphorylate phosphoinositides with a certain degree of specificity and the impairments of which are often linked to human diseases. This degree of multiplicity at the enzyme level together with the universal roles of these lipids in cell regulation assumes that inositol lipids are spatially and functionally restricted in specific membrane compartments. Studying the compartmentalized roles of these lipids at the cellular level represents a major methodological challenge. Over the last 10 years significant progress has been made in creating reagents that can monitor inositol lipid changes in live cells with fluorescence or confocal microscopy. New methods are also being developed to manipulate these lipids in specific membrane compartments in a regulated fashion. This article recalls some historical aspects of inositide research and describes the new methodological advances highlighting their great potential as well as the problems one can encounter with their use.