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Manganese

氯化锰(II)四水合物

Company: Sigma-Aldrich
Catalog#: M3634
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In vitro AMPylation/Adenylylation of Alpha-synuclein by HYPE/FICD
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 ...

Soluble and Solid Iron Reduction Assays with Desulfitobacterium hafniense
Author:
Date:
2018-09-05
[Abstract]  There is a pressing need to develop sustainable and efficient methods to protect and stabilize iron objects. To develop a conservation-restoration method for corroded iron objects, this bio-protocol presents the steps to investigate reductive dissolution of ferric iron and biogenic production of stabilizing ferrous iron minerals in the strict anaerobe Desulfitobacterium hafniense (strains TCE1 and LBE). We investigated iron reduction using three different Fe(III) sources: Fe(III)-citrate (a soluble phase), akaganeite (solid iron phase), and corroded coupons. This protocol describes a method that combines spectrophotometric quantification of the complex Fe(II)-Ferrozine® with mineral characterization by scanning electron microscopy and Raman spectroscopy. These three ... [摘要]  迫切需要开发可持续和有效的方法来保护和稳定铁制物体。为了开发腐蚀铁物体的保护 - 恢复方法,该生物方案提出了研究严格厌氧菌[Desulfitobacterium hafniense (菌株TCE1)中三价铁的还原溶解和稳定亚铁矿物质的生物产生的步骤。和LBE)。我们使用三种不同的Fe(III)来源研究了铁还原:Fe(III) - 柠檬酸盐(可溶相),akaganeite(固体铁相)和腐蚀的试样。该协议描述了一种方法,该方法结合了复杂的Fe(II)-Ferrozine ®的分光光度定量,通过扫描电子显微镜和拉曼光谱进行矿物表征。这三种方法可以评估三价铁的还原溶解和生物矿物质生产,作为开发一种创新的可持续方法来稳定腐蚀铁的有希望的替代方法。

【背景】自铁器时代以来,铁已被用于生产日常用具。因此,考古学上的铁试验是过去极其重要的证据,应予以保留。然而,由于其反应性,铁容易被腐蚀并且考古铁物体可能被完全损坏。埋藏时,铁制品会根据埋葬地点的环境条件形成复杂的腐蚀层。挖掘后,条件发生变化,腐蚀层变得不稳定。为避免完全破坏,考古铁制物需要快速稳定处理。目前,可用的稳定化处理不能提供长期保护并且具有实质性缺点,例如毒性,低效率和大量废物的产生(Scott和Eggert,2009; Rimmer 等人, 2012)。因此,有必要开发新技术来稳定考古铁器。

越来越多地考虑利用微生物代谢来开发更有效,可持续和环保的保护 ...

Nuclear Transformation of Chlamydomonas reinhardtii by Electroporation
Author:
Date:
2018-05-05
[Abstract]  The unicellular green alga Chlamydomonas reinhardtii is an important model organism for studying photosynthesis, acclimation to abiotic stress, cilia biology, and many other biological processes. Many molecular biology tools exist for interrogating gene function including the ability to easily transform the nuclear genome of Chlamydomonas. While technical advances such as TALENs, ZFNs and CRISPR are making it easier to precisely edit the nuclear genome, the efficiency of such methods in Chlamydomonas is at present very low. In contrast, random insertion by nuclear transformation tends to be a much more efficient process. This protocol describes a method for transformation of the Chlamydomonas nuclear genome by electroporation. The protocol requires at ... [摘要]  单细胞绿藻莱茵衣藻是研究光合作用,适应非生物胁迫,纤毛生物学和许多其他生物过程的重要模式生物。 许多分子生物学工具用于询问基因功能,包括轻松转化衣藻的核基因组的能力。 虽然TALENs,ZFNs和CRISPR等技术进步正在使精确编辑核基因组变得更加容易,但此类方法在衣原体中的效率目前非常低。 相反,通过核转变随机插入往往是一个更有效的过程。 该协议描述了通过电穿孔转化衣原体核基因组的方法。 该协议需要至少3天的工作,并通常导致1-2周内出现小菌落。

【背景】众多的分子,遗传和基因组资源使得莱茵衣藻(以下简称衣衣属)成为研究各种生物过程的优秀模式生物。已经开发了许多技术来改变衣藻核,叶绿体和线粒体,包括粒子轰击(Boynton等,1988),玻璃珠转化(Kindle,1990)和电穿孔(Shimogawara等人,1998)。核衣壳菌可通过将衣藻暴露于物理或化学诱变剂(例如紫外线或甲磺酸乙酯)而产生,但通常通过随机插入诱变转基因DNA而获得。由于衣藻核转化的同源重组效率非常低(Zorin等人,2009; Jinkerson和Jonikas,2015),转化的DNA通常整合到核基因组随机位点。存在许多用于随后鉴定异位DNA的插入位点的技术,包括经典遗传作图(Rymarquis等人,2005),TAIL-PCR(Dent等人 ...

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