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The Mitoribosome

Methods and Protocols

  • Book
  • © 2023

Overview

Editors:
  1. Antoni Barrientos

    1. Miller School of Medicine, University of Miami, Miami, USA

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    You can also search for this editor in PubMed   Google Scholar

  2. Flavia Fontanesi

    1. Miller School of Medicine, University of Miami, Miami, USA

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    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2661)

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About this book

This volume provides classic and new methods to study the structure, assembly pathway, and protein synthesis ability of mitoribosomes across species.  Following an introduction of fundamental concepts on the topic, method chapters present detailed protocols based on cryo-electron tomography, cryo-EM approaches, mitoribosome purification techniques, mitochondrial translation assays, and methods to study mitochondrial mRNAs that are translated on mitoribosomes. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

 

Authoritative and cutting-edge, The Mitoribosome: Methods and Protocols, aims to be a comprehensive guide for researchers in the field.

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Table of contents (19 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. General Concepts

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. Discovery of Mitochondrial Ribosomes

      • Thomas W. O’Brien
      Pages 3-5

    3. Evolution: Mitochondrial Ribosomes Across Species

      • Rajendra K. Agrawal, Soneya Majumdar
      Pages 7-21

    4. Mitoribosome Biogenesis

      • J. Conor Moran, Samuel Del’Olio, Austin Choi, Hui Zhong, Antoni Barrientos
      Pages 23-51

    5. Translation in Mitochondrial Ribosomes

      • Zofia M. Chrzanowska-Lightowlers, Robert N. Lightowlers
      Pages 53-72

  3. Methods to Study Mitoribosome Structure, Function, and Biogenesis

    1. Front Matter

      Pages 73-73
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    2. Sample Preparation of Isolated Mitochondria for Cryoelectron Tomography and In Situ Studies of Translation

      • Lena Thärichen, Robert Englmeier, Friedrich Förster
      Pages 75-88

    3. Cryo-EM for Structure Determination of Mitochondrial Ribosome Samples

      • Hauke S. Hillen
      Pages 89-100

    4. Sucrose Gradient Analysis of Human Mitochondrial Ribosomes and RNA

      • Kah Ying Ng, Brendan J. Battersby
      Pages 101-117

    5. Yeast Mitoribosome Purification and Analyses by Sucrose Density Centrifugation and Immunoprecipitation

      • Andreas Aufschnaiter, Andreas Carlström, Martin Ott
      Pages 119-132

    6. Rapid Cryopurification of the Yeast Mitochondrial Ribosome

      • Hong Weng Pang, Antoni Barrientos
      Pages 133-141

    7. Methods to Study the Biogenesis of Mitoribosomal Proteins in Yeast

      • Lea Bertgen, Tamara Flohr, Johannes M. Herrmann
      Pages 143-161

    8. Systematic Analysis of Assembly Intermediates in Yeast to Decipher the Mitoribosome Assembly Pathway

      • Samuel Del’Olio, Antoni Barrientos
      Pages 163-191

    9. Metabolic Labeling of Mitochondrial Translation Products in Whole Cells and Isolated Organelles

      • Priyanka Maiti, Flavia Fontanesi
      Pages 193-215

    10. Assembly of the Mitochondrial Translation Initiation Complex

      • Cristina Remes, Minh Duc Nguyen, Henrik Spahr, Martin Ng, Clark Fritsch, Arpan Bhattacharya et al.
      Pages 217-232

    11. Reconstitution of Mammalian Mitochondrial Translation System Capable of Long Polypeptide Synthesis

      • Muhoon Lee, Nono Takeuchi-Tomita
      Pages 233-255

    12. Human Mitoribosome Profiling: A Re-engineered Approach Tailored to Study Mitochondrial Translation

      • Iliana Soto, Mary Couvillion, L. Stirling Churchman
      Pages 257-280

    13. The ARG8m Reporter for the Study of Yeast Mitochondrial Translation

      • Daniel Flores-Mireles, Yolanda Camacho-Villasana, Xochitl Pérez-Martínez
      Pages 281-301

    14. Four-Color STED Super-Resolution RNA Fluorescent In Situ Hybridization and Immunocytochemistry to Visualize Mitochondrial mRNAs in Context with Mitochondrial Ribosomes

      • Christin A. Albus, Rolando Berlinguer-Palmini, Zofia M. Chrzanowska-Lightowlers, Robert N. Lightowlers
      Pages 303-316
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Editors and Affiliations

  • Miller School of Medicine, University of Miami, Miami, USA

    Antoni Barrientos, Flavia Fontanesi

Bibliographic Information

  • Book Title: The Mitoribosome

  • Book Subtitle: Methods and Protocols

  • Editors: Antoni Barrientos, Flavia Fontanesi

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-3171-3

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2023

  • Hardcover ISBN: 978-1-0716-3170-6Published: 12 May 2023

  • Softcover ISBN: 978-1-0716-3173-7Due: 25 May 2024

  • eBook ISBN: 978-1-0716-3171-3Published: 11 May 2023

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 346

  • Number of Illustrations: 1 b/w illustrations, 40 illustrations in colour

  • Topics: Genetics and Genomics

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