Ex Uno Plura: Differential Labeling of Phospholipid Biosynthetic Pathways with a Single Bioorthogonal Alcohol,Biochemistry

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Ex Uno Plura: Differential Labeling of Phospholipid Biosynthetic Pathways with a Single Bioorthogonal Alcohol
Biochemistry ( IF 2.9 ) Pub Date : 2017-11-08 00:00:00 , DOI: 10.1021/acs.biochem.7b01021
Timothy W. Bumpus ₁ , Felice J. Liang ₁ , Jeremy M. Baskin ₁

Affiliation

Imaging approaches that track biological molecules within cells are essential tools in modern biochemistry. Lipids are particularly challenging to visualize, as they are not directly genetically encoded. Phospholipids, the most abundant subgroup of lipids, are structurally diverse and accomplish many cellular functions, acting as major structural components of membranes and as signaling molecules that regulate cell growth, division, apoptosis, cytoskeletal dynamics, and numerous other physiological processes. Cells regulate the abundance, and therefore bioactivity, of phospholipids by modulating the activities of their biosynthetic enzymes. Thus, techniques that enable monitoring of flux through individual lipid biosynthetic pathways can provide key functional information. For example, the choline analogue propargylcholine (ProCho) can report on de novo biosynthesis of phosphatidylcholine by conversion to an alkynyl lipid that can be imaged following click chemistry tagging with an azido fluorophore. We report that ProCho is also a substrate of phospholipase D enzymes—which normally hydrolyze phosphatidylcholine to generate the lipid second messenger phosphatidic acid—in a transphosphatidylation reaction, generating the identical alkynyl lipid. By controlling the activities of phosphatidylcholine biosynthesis and phospholipase D enzymes, we establish labeling conditions that enable this single probe to selectively report on two different biosynthetic pathways. Just as nature exploits the economy of common metabolic intermediates to efficiently diversify biosynthesis, so can biochemists in interrogating such pathways with careful probe design. We envision that ProCho’s ability to report on multiple metabolic pathways will enable studies of membrane dynamics and improve our understanding of the myriad roles that lipids play in cellular homeostasis.

中文翻译：

Ex Uno Plura：单一生物正交酒精对磷脂生物合成途径的差异标记

追踪细胞内生物分子的成像方法是现代生物化学中必不可少的工具。脂质在可视化方面特别具有挑战性，因为它们不是直接遗传编码的。磷脂是脂质中最丰富的亚组，其结构多样，可以完成许多细胞功能，它们充当膜的主要结构成分，并作为调节细胞生长，分裂，凋亡，细胞骨架动力学和许多其他生理过程的信号分子。细胞通过调节其生物合成酶的活性来调节磷脂的丰度，从而调节其生物活性。因此，能够通过单个脂质生物合成途径监测通量的技术可以提供关键的功能信息。例如，胆碱类似物炔丙基胆碱（ProCho）可以报告通过重新合成为炔基脂质可以重新合成磷脂酰胆碱的生物合成方法，在点击化学标记上叠氮基荧光团后即可成像。我们报道ProCho还是磷脂酶D酶的底物，后者通常在转磷脂酰化反应中水解磷脂酰胆碱以生成脂质第二信使磷脂酸，从而生成相同的炔基脂质。通过控制磷脂酰胆碱生物合成和磷脂酶D酶的活性，我们建立了标记条件，使该单个探针能够选择性地报告两种不同的生物合成途径。正如大自然利用常见的代谢中间体的经济性来有效地实现生物合成的多样化一样，生物化学家也可以通过仔细的探针设计来探究这些途径。我们设想ProCho报告多种代谢途径的能力将使人们能够研究膜动力学，并加深我们对脂质在细胞稳态中所起的多种作用的理解。

更新日期：2017-11-09

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