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Author:
Date:
2020-12-05
[Abstract] Pipecolic acid (Pip), a non-proteinacious product of lysine catabolism, is an important regulator of immunity in plants and humans alike. For instance, Pip accumulation is associated with the genetic disorder Zellweger syndrome, chronic liver diseases, and pyridoxine-dependent epilepsy in humans. In plants, Pip accumulates upon pathogen infection and is required for plant defense. The aminotransferase ALD1 catalyzes biosynthesis of Pip precursor piperideine-2-carboxylic acid, which is converted to Pip via ornithine cyclodeaminase. A variety of methods are used to quantify Pip, and some of these involve use of expensive amino acid analysis kits. Here, we describe a simplified procedure for quantitative analysis of Pip from plant tissues. Pipecolic acid was extracted from leaf tissues along ...
[摘要] [摘要]胡椒酸（Pip）是赖氨酸分解代谢的非蛋白质产物，是植物和人类免疫力的重要调节剂。例如，Pip积累与人类遗传病Zellweger综合征，慢性肝病和吡ido醇依赖性癫痫有关。在植物中，Pip会在病原体感染时积聚，是植物防御所必需的。氨基转移酶ALD1催化Pip前体哌啶-2-羧酸的生物合成，后者通过鸟氨酸环脱氨酶转化为Pip。多种方法可用于定量Pip，其中一些方法涉及使用昂贵的氨基酸分析试剂盒。在这里，我们描述了从植物组织中定量定量分析的简化程序。P ipecolic酸是从叶组织中提取与内标正缬氨酸一起衍生与丙基氯甲酸酯和分析通过气相色谱-质谱联用选择性离子模式。此过程简单，经济且有效，并且不涉及同位素内标或多步衍生化。
关键词：胡椒酸GC-MS拟南芥植物防御氯甲酸丙酯
[背景]匹，非蛋白氨基酸赖氨酸的氨基酸衍生物，积聚在病原体感染的植物的本地和远端的组织和患者的体液与过氧化物酶体疾病（Schutgens等人，1986;尹和An，2010;Návarová等人。，2012;王等人，2018） ...
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Author:
Date:
2020-09-20
[Abstract] One of the major histopathological hallmarks of Parkinson’s disease are Lewy bodies (LBs) –cytoplasmic inclusions, enriched with fibrillar forms of the presynaptic protein alpha-synuclein (α-syn). Progressive deposition of α-syn into LBs is enabled by its propensity to fibrillize into insoluble aggregates. We recently described a marked reduction in α-syn fibrillation in vitro upon posttranslational modification (PTM) by the Fic (Filamentation induced by cAMP) family adenylyltransferase HYPE/FICD (Huntingtin yeast-interacting protein E/FICD). Specifically, HYPE utilizes ATP to covalently decorate key threonine residues in α-syn’s N-terminal and NAC (non-amyloid-β component) regions with AMP (adenosine monophosphate), in a PTM termed AMPylation or adenylylation. Status quo in ...
[摘要] [摘要 ] 帕金森氏病的主要组织病理学标志之一是路易体（LB） –细胞质内含物，富含纤维状形式的突触前蛋白α-突触核蛋白（α-syn）。由于α-syn易于原纤维化成不溶性聚集体，因此可以逐步沉积到LBs中。我们最近描述了Fic（由cAMP诱导的细丝化）家族腺苷酸转移酶HYPE / FICD（与亨廷顿酵母相互作用的蛋白E / FICD）在翻译后修饰（PTM）后体外α-syn纤颤的明显减少。具体而言，HYPE利用ATP在称为AMPylation 或adenylylation 的PTM中，以AMP（单磷酸腺苷）共价修饰α-syn's N末端和NAC（非淀粉样β成分）区域中的关键苏氨酸残基。HYPE底物（例如α-syn）的体外AMPyl化反应现状使用多种ATP类似物，包括放射性标记的α - 32 P-ATP 或α - 33 P-ATP，荧光ATP类似物，生物素化ATP类似物（N6- [6-六甲基] -ATP-生物素），以及基于点击化学的烷基-ATP方法检测基于凝胶的AMPylation 。当前描述HYPE介导AMPylation 的分步方案的文献依赖于α - 33 P-ATP核苷酸，而不是更常见的α - 32 P-ATP。尽管有效，但前一种方法需要长时间且危险的DMSO-PPO（二甲基亚砜- 聚苯基恶唑）沉淀。因此，我们提供了基于α - 33 ...
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