生物化学

分类

    现刊
    In vitro Nitrate Reductase Activity Assay of Mycolicibacterium smegmatis Crude Extract
    耻垢分枝杆菌粗提物硝酸还原酶活性的体外测定
    作者:Wei Tan, Zhi-Hui Shao and Guo-Ping Zhao日期:07/20/2021,浏览量:359,Q&A: 0
    [Abstract]

    Nitrate is one of the major inorganic nitrogen sources for microorganisms. Many bacterial and archaeal lineages can express cytoplasmic assimilatory nitrate reductase (NAS), which catalyzes the rate-limiting reduction of nitrate to nitrite in the nitrate assimilation pathway. Here, we present a detailed protocol for measuring in vitro nitrate

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    Characterising Plant Deubiquitinases with in vitro Activity-based Labelling and Ubiquitin Chain Disassembly Assays
    植物去泛素酶的体外活性标记和泛素链分解分析
    作者:Michael J. Skelly and Steven H. Spoel日期:05/05/2021,浏览量:2033,Q&A: 0
    [Abstract]

    Post-translational modification of proteins by ubiquitin is an essential cellular signaling mechanism in all eukaryotes. Ubiquitin is removed from target proteins by a wide range of deubiquitinase (DUB) enzymes with different activities and substrate specificities. Understanding how DUBs function in vitro is a vital first step to uncovering their

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    In vitro Reconstitution Assays of Arabidopsis 20S Proteasome
    拟南芥20S蛋白酶体的体外重组分析
    作者:Yanjun Li, Di Sun, Xingxing Yan, Zhiye Wang and Xiuren Zhang日期:04/05/2021,浏览量:2794,Q&A: 0
    [Abstract]

    The majority of cellular proteins are degraded by the 26S proteasome in eukaryotes. However, intrinsically disordered proteins (IDPs), which contain large portions of unstructured regions and are inherently unstable, are degraded via the ubiquitin-independent 20S proteasome. Emerging evidence indicates that plant IDP homeostasis may also be

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    Quantitative Measurement of Mucolytic Enzymes in Fecal Samples
    粪便粘液溶解酶的定量测定
    作者:Shahanshah Khan and Hasan Zaki日期:03/20/2021,浏览量:1918,Q&A: 0
    [Abstract]

    The mucus layer in the gastrointestinal tract covers the apical surface of intestinal epithelial cells, protecting the mucosal tissue from enteric pathogen and commensal microorganisms. The mucus is primarily composed of glycosylated protein called mucins, which are produced by goblet cells, a type of columnar epithelial cells in the intestinal

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    Polyamine Transport Assay Using Reconstituted Yeast Membranes
    利用重构酵母细胞膜的多胺转运实验
    作者:Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe日期:01/20/2021,浏览量:1382,Q&A: 0
    [Abstract]

    ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure

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    Assessment of Diadenylate Cyclase and c-di-AMP-phosphodiesterase Activities Using Thin-layer and Ion Exchange Chromatography
    使用薄层和离子交换色谱评估二腺苷酸环化酶和c-di-AMP-磷酸二酯酶的活性
    作者:Andreas Latoscha, David Jan Drexler, Gregor Witte and Natalia Tschowri日期:01/05/2021,浏览量:1021,Q&A: 0
    [Abstract]

    All living cells use cyclic nucleotides as second messengers for signal sensing and transduction. Cyclic di-3′,5′-adenosine monophosphate (c-di-AMP) is primarily involved in the control of bacterial and euryarcheal osmoadaptation and is produced by diadenylate cyclases from two molecules of ATP. Specific phosphodiesterases hydrolyze

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    Non-radioactive Assay to Determine Product Profile of Short-chain Isoprenyl Diphosphate Synthases
    用于确定短链异戊烯基二磷酸合酶产物谱的非放射性分析
    作者:Avanish Rai and Dinesh A. Nagegowda日期:01/05/2021,浏览量:1033,Q&A: 0
    [Abstract] Isoprenoids represent the largest class of metabolites with amazing diversities in structure and function. They are involved in protecting plants against pathogens or herbivores or involved in attracting pollinators. Isoprenoids are derived from geranyl diphosphate (GPP; C10), farnesyl diphosphate (FPP; C15), geranylgeranyl ...
    The ATPase Activity of Escherichia coli Expressed AAA+-ATPase Protein
    大肠杆菌表达的AAA + -ATPase蛋白的ATPase活性
    作者:Amita Kaundal, Vemanna S. Ramu and Kirankumar S. Mysore日期:08/05/2020,浏览量:1810,Q&A: 0
    [Abstract] ATPases are the enzymes that breakdown ATP to ADP and release inorganic phosphate (Pi). Here we provide a detailed protocol to determine the ATPase activity of a recombinant AAA+-ATPase protein (GENERAL CONTROL NON-REPRESSIBLE-4 [GCN4]) by spectrophotometric absorption at 360 nm to measure the accumulated inorganic phosphate. In ...
    Quantification of Protein Kinase A (PKA) Activity by An in vitro Radioactive Assay Using the Mouse Sperm Derived Enzyme
    用小鼠精子衍生酶体外放射性测定蛋白激酶A(PKA)活性
    作者:Cintia Stival, Carolina Baro Graf, Pablo E. Visconti and Dario Krapf日期:06/20/2020,浏览量:2133,Q&A: 0
    [Abstract] In order to acquire fertilizing potential, mammalian sperm must undergo a process known as capacitation, which relies on the early activation of Protein Kinase A (PKA). Frequently, PKA activity is assessed in whole-cell experiments by analyzing the phosphorylation status of its substrates in a western-blot. This technique ...
    Preparation of HeLa Total Membranes and Assay of Lipid-inhibition of Serine Palmitoyltransferase Activity
    HeLa全膜的制备及其对丝氨酸-棕榈酰转移酶活性的抑制作用
    作者:Muthukumar Kannan, Deanna L. Davis, John Suemitsu, Christopher D. Oltorik and Binks Wattenberg日期:06/20/2020,浏览量:1716,Q&A: 0
    [Abstract] Serine palmitoyltranferase (SPT) is a pyridoxal 5′ phosphate (PLP)-dependent enzyme that catalyzes the first and rate-limiting step of de novo synthesis of sphingolipids. SPT activity is homeostatically regulated in response to increased levels of sphingolipids. This homeostatic regulation of SPT is mediated through small ER membrane ...