Biochemistry

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    Protocols in Current Issue
    Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids
    Authors:  Shunshi Kohyama, Kei Fujiwara, Natsuhiko Yoshinaga and Nobuhide Doi, date: 03/20/2020, view: 171, Q&A: 0
    [Abstract] The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling ...
    In vitro Measurement of CMP-Sialic Acid Transporter Activity in Reconstituted Proteoliposomes
    Authors:  James Cahill, Shivani Ahuja and Matthew R. Whorton, date: 03/20/2020, view: 223, Q&A: 0
    [Abstract] Nucleotide-sugar transporters (NSTs) facilitate eukaryotic cellular glycosylation by transporting nucleotide-sugar conjugates into the Golgi lumen and endoplasmic reticulum for use by glycosyltransferases, while also transferring nucleotide monophosphate byproducts to the cytoplasm. Mutations in this family of proteins can cause a number of ...
    Contemporaneous Measurement of Outer and Inner Membrane Permeability in Gram-negative Bacteria
    Authors:  Bo Ma, Chao Fang, Jing Zhang, Mingzhi Wang, Xiaoxing Luo and Zheng Hou, date: 03/05/2020, view: 260, Q&A: 0
    [Abstract] The emergence and rapid spread of multidrug resistance in bacteria have led to the urgent need for novel antibacterial agents. Membrane permeabilization is the mechanism for many antibacterial molecules that are being developed against gram-negative bacteria. Thus, to determine the efficacy of a potential antibacterial molecule, it is important to ...
    Lipid Mixing Assay for Murine Myoblast Fusion and Other Slow Cell-cell Fusion Processes
    [Abstract] Lipid mixing (redistribution of lipid probes between fusing membranes) has been widely used to study early stages of relatively fast viral and intracellular fusion processes that take seconds to minutes. Lipid mixing assays are especially important for identification of hemifusion intermediates operationally defined as lipid mixing without content ...
    Photoactivable Cholesterol as a Tool to Study Interaction of Influenza Virus Hemagglutinin with Cholesterol
    Authors:  Bodan Hu, Mohamed Rasheed Gadalla, Christoph Thiele and Michael Veit, date: 02/20/2020, view: 260, Q&A: 0
    [Abstract] Non-covalent binding of cholesterol to the transmembrane region of proteins affect their functionalities, but methods to prove such an interaction are rare. We describe our protocol to label the hemagglutinin (HA) of Influenza virus with a cholesterol derivative in living cells or with immunoprecipitated protein. We synthesized a “clickable” ...
    Eicosanoid Isolation from Mouse Intestinal Tissue for ELISA
    Author:  Isabella Rauch, date: 11/05/2018, view: 1786, Q&A: 0
    [Abstract] Activation of inflammasomes in peritoneal macrophages and intestinal epithelial cells (IEC) leads to the release of eicosanoids. To assess the amount of eicosanoids released by IEC, lipids need to be isolated from whole tissue previous to analysis by lipid mass spectrometry or ELISA. This protocol describes how to isolate lipids from intestinal ...
    GC-MS-Based Analysis of Methanol: Chloroform-extracted Fatty Acids from Plant Tissues
    Authors:  Manish Kumar Patel, Shubhashis Das and Jitendra Kumar Thakur, date: 09/20/2018, view: 5603, Q&A: 0
    [Abstract] Fatty acids (FAs) are carboxylic acids with long aliphatic chains that may be straight, branched and saturated or unsaturated. Most of the naturally occurring plant FAs contains an even number of carbon (C4-C24). FAs are used in food and pharmacological industries due to their nutritional importance. In addition, FAs are considered as a promising ...
    Liposome Flotation Assay for Studying Interactions Between Rubella Virus Particles and Lipid Membranes
    Authors:  Kyoko Saito, Noriyuki Otsuki, Makoto Takeda and Kentaro Hanada, date: 08/20/2018, view: 3751, Q&A: 0
    [Abstract] Rubella virus (RuV) is an enveloped, positive-sense single-stranded RNA virus that is pathogenic to humans. RuV binds to the target cell via the viral envelope protein E1, but the specific receptor molecules on the target cell are yet to be fully elucidated. Here, we describe a protocol for liposome flotation assay to study direct interactions ...
    Modifying Styrene-maleic Acid Co-polymer for Studying Lipid Nanodiscs by Direct Fluorescent Labeling
    Authors:  Victoria Schmidt and James N. Sturgis, date: 08/20/2018, view: 2938, Q&A: 0
    [Abstract] This protocol was developed to functionalize styrene maleic acid (SMA) by direct fluorescent labeling in an easy way, accessible to biochemistry laboratories. This novel method is based on the coupling of carboxylic acids to primary amines using a carbodiimide, a reaction commonly used for protein chemistry. The procedure uses the hydrolyzed ...
    Assessment of Uptake and Biodistribution of Radiolabeled Cholesterol in Mice Using Gavaged Recombinant Triglyceride-rich Lipoprotein Particles (rTRL)
    Authors:  Anna Worthmann, Clara John and Joerg Heeren, date: 07/05/2018, view: 2559, Q&A: 0
    [Abstract] The here described method can be used to estimate the uptake of orally provided cholesterol in mice. Briefly, mice are gavaged with radiolabeled cholesterol and 4 h later, organ distribution of the radiolabel is determined by liquid scintillation counting. The method has been applied successfully to determine dietary cholesterol handling of mice ...



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