Biochemistry

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    Protocols in Current Issue
    Imaging Microtubules in vitro at High Resolution while Preserving their Structure
    Authors:  Camille Cuveillier, Yasmina Saoudi, Isabelle Arnal and Christian Delphin, date: 04/05/2021, view: 542, Q&A: 0
    [Abstract]

    Microtubules (MT) are the most rigid component of the cytoskeleton. Nevertheless, they often appear highly curved in the cellular context and the mechanisms governing their overall shape are poorly understood. Currently, in vitro microtubule analysis relies primarily on electron microscopy for its high resolution and Total Internal Reflection

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    Simultaneous Imaging of Single Protein Size, Charge, and Binding Using A Protein Oscillation Approach
    Authors:  Guangzhong Ma, Zijian Wan and Shaopeng Wang, date: 03/05/2021, view: 1550, Q&A: 0
    [Abstract]

    Electrophoresis and Western blot are important tools in protein research for detection and identification of proteins. These traditional techniques separate the proteins based on size and charge differences and identify the proteins by antibody binding. Over the past decade, the emergence of single-molecule techniques has shown great potential in

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    Live Cell FRET Analysis of the Conformational Changes of Human P-glycoprotein
    Authors:  Ryota Futamata, Noriyuki Kioka and Kazumitsu Ueda, date: 02/20/2021, view: 1411, Q&A: 0
    [Abstract]

    The molecular mechanisms of P-glycoprotein (P-gp; also known as MDR1 or ABCB1) have been mainly investigated using artificial membranes such as lipid-detergent mixed micelles, artificial lipid bilayers, and membrane vesicles derived from cultured cells. Although these in vitro experiments help illustrate details about the molecular mechanisms of

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    Determination of Microtubule Lattice Parameters from Cryo-electron Microscope Images Using TubuleJ
    Authors:  Siou Ku, Cédric Messaoudi, Charlotte Guyomar, Charles Kervrann and Denis Chrétien, date: 11/05/2020, view: 1112, Q&A: 0
    [Abstract]

    The α-β tubulin heterodimer undergoes subtle conformational changes during microtubule assembly. These can be modulated by external factors, whose effects on microtubule structure can be characterized on 2D views obtained by cryo-electron microscopy. Analysis of microtubule images is facilitated if they are straight enough to interpret

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    Protocols for Processing and Interpreting cryoEM Data Using Bsoft: A Case Study of the Retinal Adhesion Protein, Retinoschisin
    Author:  J. Bernard Heymann, date: 01/20/2020, view: 2397, Q&A: 0
    [Abstract] The goal of cryoEM is to determine the structures of biomolecules from electron micrographs. In many cases the processing is straightforward and can be handled with routine protocols. In other cases, the properties and behavior of the specimen require adaptions to properly interpret the data. Here I describe the protocols for examining the higher ...
    Isolation and Imaging of His- and RFP-tagged Amyloid-like Proteins from Caenorhabditis elegans by TEM and SIM
    Authors:  Amberley D. Stephens, Meng Lu and Gabriele S. Kaminski Schierle, date: 11/05/2019, view: 2172, Q&A: 0
    [Abstract] In our recently published paper, we highlight that during normal aging of C. elegans age-dependent aggregates of proteins form and lead to functional decline of tissues. The protocol described here details the isolation of two proteins from C. elegans in their aggregated amyloid-like form, casein kinase I isoform alpha (KIN-19) ...



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