Investigating The Structure Of Macromolecular Complexes And Membrane Proteins Using Transmission Electron Microscopy

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Investigating the Structure of Macromolecular Complexes and Membrane Proteins Using Transmission Electron Microscopy

Author : Breanna Sachiko Vollmar
Publisher :
ISBN 13 :
Total Pages : 252 pages
Book Rating : 4.:/5 (879 download)

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Book Synopsis Investigating the Structure of Macromolecular Complexes and Membrane Proteins Using Transmission Electron Microscopy by : Breanna Sachiko Vollmar

Download or read book Investigating the Structure of Macromolecular Complexes and Membrane Proteins Using Transmission Electron Microscopy written by Breanna Sachiko Vollmar and published by . This book was released on 2013 with total page 252 pages. Available in PDF, EPUB and Kindle. Book excerpt: Transmission electron microscopy (TEM) has provided atomic resolution details regarding the biological processes that give rise to life. The work presented here focuses on the application of TEM for studying membrane proteins and macromolecular assemblies using electron crystallography and single particle electron cryomicroscopy (cryo-EM). Two membrane proteins, human aquaporin 3 (hAQP3) and galactose permease (GalP) from Escherichia coli, were studied using electron crystallography. hAQP3 was successfully expressed in Pichia pastoris and purified, but failed to reconstitute to form two-dimensional crystals. Two-dimensional crystals of GalP were optimized to form large, crystalline vesicles and sheets. A projection map of GalP calculated from a cryo-EM image showed that GalP forms trimers in the two-dimensional crystals. Single particle TEM studies focused on three macromolecular assemblies, alphaB-crystallin, a designed octahedral protein cage and a mutant HCV IRES-40S ribosome complex. Projection class averages of negatively stained alphaB-crystallin were combined with NMR and SAXS data to produce a pseudo-atomic model of alphaB-crystallin and provide a hypothesis for multimerization. A single particle cryo-EM reconstruction of a designed self-assembling octahedral protein oligomer showed that a designed protein assembly forms the predicted symmetry in solution. A 17 [angstrom] resolution single particle cryo-EM reconstruction of a mutant HCV IRES bound to the 40S ribosomal subunit revealed that deletion of three nucleotides in the IRES modulate interaction with the 40S ribosomal subunit. The conformational changes induced in the 40S were shown to affect a step after 80S formation, the transition from initiation to elongation. These studies illustrate the application of TEM to study the molecular mechanisms used by both membrane proteins and macromolecular complexes to carry out biological processes.

Three-Dimensional Electron Microscopy of Macromolecular Assemblies

Author : Frank Joachim
Publisher : Elsevier
ISBN 13 : 0080525814
Total Pages : 361 pages
Book Rating : 4.0/5 (85 download)

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Book Synopsis Three-Dimensional Electron Microscopy of Macromolecular Assemblies by : Frank Joachim

Download or read book Three-Dimensional Electron Microscopy of Macromolecular Assemblies written by Frank Joachim and published by Elsevier. This book was released on 1996-01-24 with total page 361 pages. Available in PDF, EPUB and Kindle. Book excerpt: Three-Dimensional Electron Microscopy of Macromolecular Assemblies is the first systematic introduction to single-particle methods of reconstruction. It covers correlation alignment, classification, 3D reconstruction, restoration, and interpretation of the resulting 3D images in macromolecular assemblies. It will be an indispensable resource for newcomers to the field and for all using or adopting these methods. Key Features* Presents methods that offer an alternative to crystallographic techniques for molecules that cannot be crystallized* Describes methods that have been instrumental in exploring the three-dimensional structure of* the nuclear pore complex* the calcium release channel;* the ribosome* chaperonins

Protein Structure Determination on Lipid Substrates by Electron Microscopy

Author : Elizabeth Morrison Wilson-Kubalek
Publisher :
ISBN 13 : 9780542002434
Total Pages : 344 pages
Book Rating : 4.0/5 (24 download)

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Book Synopsis Protein Structure Determination on Lipid Substrates by Electron Microscopy by : Elizabeth Morrison Wilson-Kubalek

Download or read book Protein Structure Determination on Lipid Substrates by Electron Microscopy written by Elizabeth Morrison Wilson-Kubalek and published by . This book was released on 2005 with total page 344 pages. Available in PDF, EPUB and Kindle. Book excerpt: With the impressive growth in gene sequence data and advances in genetics the number of proteins and macromolecular assemblies for structural studies are increasing at a rapid rate. Structural biology will help to unravel the function of the newly discovered proteins and their molecular interactions within the cell and will ultimately provide the framework for developing new therapeutic drugs. Electron crystallography has become a valuable method for the structure determination of proteins and macromolecular complexes that are large or heterogeneous, difficult to obtain, unstable, insoluble, or that fail to yield crystals suitable for X-ray crystallographic analysis. Although electron microscopy (EM) and image analysis methods are rapidly improving and continuing to prove their value, advances in sample preparation have been limited, and general methods for obtaining ordered arrays of proteins for EM analysis are needed. In this thesis, I have described a method for promoting the formation of helical protein arrays. Helical crystallization on lipid nanotubes presents an alternative platform to explore fundamentals of protein ordering, intermolecular protein interaction and phase behavior. Three-dimensional (3-D) maps of wild-type and mutant streptavidin (SA) show that the molecular packing arrangements of SA on the surface of the lipid nanotubes differ from previously reported lattice packing of SA on biotinylated monolayers. His-tagged Perfringolysin O (PFO), a soluble toxin was used to demonstrate the utility of nickel-lipid nanotubes as a tool for structural studies of His-tagged proteins. Cryo-EM and image analysis of PFO helical arrays were used to reconstruct a 3-D density map. These studies have demonstrated that lipid nanotubes offer a robust and reproducible substrate for forming helical protein arrays which present a means for studying protein structure and structure-function relationships. Using EM and single-particle image analysis I calculated 2-D projection maps from images of oligomeric PFO prepore and pore complexes formed on lipid layers and identified an apparent conformational change from the prepore to the pore. In addition, I identified specific domains of the toxin molecule within the image and have thus determined the orientation of PFO monomers within the oligomeric complex.

Structural Investigation of Membrane Proteins by Electron Microscopy

Author : Katarzyna Beata Mościcka
Publisher :
ISBN 13 : 9789036737432
Total Pages : 148 pages
Book Rating : 4.7/5 (374 download)

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Book Synopsis Structural Investigation of Membrane Proteins by Electron Microscopy by : Katarzyna Beata Mościcka

Download or read book Structural Investigation of Membrane Proteins by Electron Microscopy written by Katarzyna Beata Mościcka and published by . This book was released on 2009 with total page 148 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Molecular Biology of The Cell

Author : Bruce Alberts
Publisher :
ISBN 13 : 9780815332183
Total Pages : 0 pages
Book Rating : 4.3/5 (321 download)

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Book Synopsis Molecular Biology of The Cell by : Bruce Alberts

Download or read book Molecular Biology of The Cell written by Bruce Alberts and published by . This book was released on 2002 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Electron Microscopy Methods and Protocols

Author : M. A. Nasser Hajibagheri
Publisher : Springer Science & Business Media
ISBN 13 : 1592592015
Total Pages : 292 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Electron Microscopy Methods and Protocols by : M. A. Nasser Hajibagheri

Download or read book Electron Microscopy Methods and Protocols written by M. A. Nasser Hajibagheri and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 292 pages. Available in PDF, EPUB and Kindle. Book excerpt: Electron Microscopy Methods and Protocols is designed for the established researcher as a manual for extending knowledge of the field. It is also for the newcomer who wishes to move into the field. A wide range of applications for the examination of cells, tissues, biological macromolecules, molecular structures, and their interactions are discussed. We have tried to gather together methods that we consider to be those most generally appli- ble to current research in both cell and molecular biology. Each chapter c- tains a set of related practical protocols with examples provided by experts who have first-hand knowledge of the techniques they describe. The individual chapters are grouped according to similarities in their specimen preparation and methodology. Methods are presented in detail, in a step-by-step fashion, using reproducible protocols the authors have personally checked. During the last decade, the scientific literature describing the use of colloidal gold as an immunocytochemical marker has increased at an ex- nential rate, and this trend is expected to continue. We have included a large number of variations on the immunogold labeling technique. In both the ne- tive staining and cryo chapters, authors emphasize the “immunological app- cations” in order to correlate as fully as possible with the emphasis on immunogold labeling in the other chapters. Electron Microscopy Methods and Protocols commences with the routine preparation of biological material for classical transmission electron microscopy involving tissue fixation, embedding, and sectioning (Chap. 1).

Structural Investigation of Macromolecular Complexes by Electron Microscopy

Author : Meikel Diepholz
Publisher :
ISBN 13 :
Total Pages : 304 pages
Book Rating : 4.:/5 (246 download)

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Book Synopsis Structural Investigation of Macromolecular Complexes by Electron Microscopy by : Meikel Diepholz

Download or read book Structural Investigation of Macromolecular Complexes by Electron Microscopy written by Meikel Diepholz and published by . This book was released on 2007 with total page 304 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Virus Protein and Nucleoprotein Complexes

Author : J. Robin Harris
Publisher : Springer
ISBN 13 : 9811084564
Total Pages : 443 pages
Book Rating : 4.8/5 (11 download)

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Book Synopsis Virus Protein and Nucleoprotein Complexes by : J. Robin Harris

Download or read book Virus Protein and Nucleoprotein Complexes written by J. Robin Harris and published by Springer. This book was released on 2018-06-13 with total page 443 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Subcellular Biochemistry series has recently embarked upon an almost encyclopaedic coverage of topics relating to the structure and function of macromolecular complexes (Volumes 82, 83 and 87). The present multi-author text covers numerous aspects of current research into molecular virology, with emphasis upon viral protein and nucleoprotein structure and function. Structural data from cryo-electron microscopy and X-ray crystallography is displayed throughout the book. The 17 chapters in the book cover diverse interesting topics, all currently under investigation, contributed by authors who are active actively involved in present-day research. Whilst structural aspects predominate, there is much consideration of the structure-function relationship. In addition, the book correlates with and extends from Volume 68 of the series “Structure and Physics of Viruses: An Integrated Textbook”. This book is directed primarily at professionals that work in the broad field of Structural Biology and will be of particular interest to Structural Virologists. The editors, David Bhella and Robin Harris, have much experience in virology and protein structure, respectively. Dr Bhella is Director of the Scottish Macromolecular Imaging Centre. Professor Robin Harris is the long-standing Series Editor of the Subcellular Biochemistry series. He has edited and contributed to several books in the series.

Three-Dimensional Electron Microscopy

Author :
Publisher : Academic Press
ISBN 13 : 0128170190
Total Pages : 303 pages
Book Rating : 4.1/5 (281 download)

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Book Synopsis Three-Dimensional Electron Microscopy by :

Download or read book Three-Dimensional Electron Microscopy written by and published by Academic Press. This book was released on 2019-07-18 with total page 303 pages. Available in PDF, EPUB and Kindle. Book excerpt: Three-Dimensional Electron Microscopy, Volume 152 in the Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters focusing on FIB-SEM of mouse nervous tissue: fast and slow sample preparation, Serial-section electron microscopy using ATUM - Automated Tape collecting Ultra-Microtome, Software for automated acquisition of electron tomography tilt series, Scanning electron tomography of biological samples embedded in plastic, Cryo-STEM tomography for Biology, CryoCARE: Content-aware denoising of cryo-EM images and tomograms using artificial neural networks, Expedited large-volume 3-D SEM workflows for comparative vertebrate microanatomical imaging, and many other interesting topics. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Cell Biology series Includes the latest information on the Three-Dimensional Electron Microscopy technique

Protein Conformational Dynamics

Author : Ke-li Han
Publisher : Springer Science & Business Media
ISBN 13 : 3319029703
Total Pages : 488 pages
Book Rating : 4.3/5 (19 download)

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Book Synopsis Protein Conformational Dynamics by : Ke-li Han

Download or read book Protein Conformational Dynamics written by Ke-li Han and published by Springer Science & Business Media. This book was released on 2014-01-20 with total page 488 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book discusses how biological molecules exert their function and regulate biological processes, with a clear focus on how conformational dynamics of proteins are critical in this respect. In the last decade, the advancements in computational biology, nuclear magnetic resonance including paramagnetic relaxation enhancement, and fluorescence-based ensemble/single-molecule techniques have shown that biological molecules (proteins, DNAs and RNAs) fluctuate under equilibrium conditions. The conformational and energetic spaces that these fluctuations explore likely contain active conformations that are critical for their function. More interestingly, these fluctuations can respond actively to external cues, which introduces layers of tight regulation on the biological processes that they dictate. A growing number of studies have suggested that conformational dynamics of proteins govern their role in regulating biological functions, examples of this regulation can be found in signal transduction, molecular recognition, apoptosis, protein / ion / other molecules translocation and gene expression. On the experimental side, the technical advances have offered deep insights into the conformational motions of a number of proteins. These studies greatly enrich our knowledge of the interplay between structure and function. On the theoretical side, novel approaches and detailed computational simulations have provided powerful tools in the study of enzyme catalysis, protein / drug design, protein / ion / other molecule translocation and protein folding/aggregation, to name but a few. This work contains detailed information, not only on the conformational motions of biological systems, but also on the potential governing forces of conformational dynamics (transient interactions, chemical and physical origins, thermodynamic properties). New developments in computational simulations will greatly enhance our understanding of how these molecules function in various biological events.

Structural Studies of Membrane Proteins Using Transmission Electron Microscopy

Author :
Publisher :
ISBN 13 : 9789175954684
Total Pages : 55 pages
Book Rating : 4.9/5 (546 download)

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Book Synopsis Structural Studies of Membrane Proteins Using Transmission Electron Microscopy by :

Download or read book Structural Studies of Membrane Proteins Using Transmission Electron Microscopy written by and published by . This book was released on 2015 with total page 55 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Membrane Protein Crystallization

Author :
Publisher : Academic Press
ISBN 13 : 0080961592
Total Pages : 334 pages
Book Rating : 4.0/5 (89 download)

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Book Synopsis Membrane Protein Crystallization by :

Download or read book Membrane Protein Crystallization written by and published by Academic Press. This book was released on 2009-05-29 with total page 334 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume of Current Topics in Membranes focuses on Membrane Protein Crystallization, beginning with a review of past successes and general trends, then further discussing challenges of mebranes protein crystallization, cell free production of membrane proteins and novel lipids for membrane protein crystallization. This publication also includes tools to enchance membrane protein crystallization, technique advancements, and crystallization strategies used for photosystem I and its complexes, establishing Membrane Protein Crystallization as a needed, practical reference for researchers.

Membrane Proteins in Aqueous Solutions

Author : JEAN-LUC. POPOT
Publisher : Springer
ISBN 13 : 9783030103248
Total Pages : 738 pages
Book Rating : 4.1/5 (32 download)

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Book Synopsis Membrane Proteins in Aqueous Solutions by : JEAN-LUC. POPOT

Download or read book Membrane Proteins in Aqueous Solutions written by JEAN-LUC. POPOT and published by Springer. This book was released on 2019-07-05 with total page 738 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is the first to be entirely devoted to the challenging art of handling membrane proteins out of their natural environment, a key process in biological and pharmaceutical research, but one plagued with difficulties and pitfalls. Written by one of the foremost experts in the field, Membrane Proteins in Aqueous Solutions is accessible to any member of a membrane biology laboratory. After presenting the structure, functions, dynamics, synthesis, natural environment and lipid interactions of membrane proteins, the author discusses the principles of extracting them with detergents, the mechanisms of detergent-induced destabilization, countermeasures, and recent progress in developing detergents with weaker denaturing properties. Non-conventional alternatives to detergents, including bicelles, nanodiscs, amphipathic peptides, fluorinated surfactants and amphipols, are described, and their relative advantages and drawbacks are compared. The synthesis and solution properties of the various types of amphipols are presented, as well as the formation and properties of membrane protein/amphipol complexes and the transfer of amphipol-trapped proteins to detergents, nanodiscs, lipidic mesophases, or living cells. The final chapters of the book deal with applications: membrane protein in vitro folding and cell-free expression, solution studies, NMR, crystallography, electron microscopy, mass spectrometry, amphipol-mediated immobilization of membrane proteins, and biomedical applications. Important features of the book include introductory sections describing foundations as well as the state-of-the-art for each of the biophysical techniques discussed, and topical tables which organize a widely dispersed literature. Boxes and annexes throughout the book explain technical aspects, and twelve detailed experimental protocols, ranging from in vitro folding of membrane proteins to single-particle electron cryomicroscopy, have been contributed by and commented on by experienced users. Membrane Proteins in Aqueous Solutions offers a concise, accessible introduction to membrane protein biochemistry and biophysics, as well as comprehensive coverage of the properties and uses of conventional and non-conventional surfactants. It will be useful both in basic and applied research laboratories and as a teaching aid for students, instructors, researchers, and professionals within the field.

Protein Structure Determination from Cryo-electron Microscopy Density Maps

Author : Yu-Ruei Wang
Publisher :
ISBN 13 :
Total Pages : 71 pages
Book Rating : 4.:/5 (932 download)

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Book Synopsis Protein Structure Determination from Cryo-electron Microscopy Density Maps by : Yu-Ruei Wang

Download or read book Protein Structure Determination from Cryo-electron Microscopy Density Maps written by Yu-Ruei Wang and published by . This book was released on 2015 with total page 71 pages. Available in PDF, EPUB and Kindle. Book excerpt: Single-particle cryo-electron microscopy (cryo-EM) has emerged as a powerful tool in structure determination of macromolecular complexes that are not suitable for crystallographic studies. Recent advances in direct electron detectors and image-processing techniques have allowed cryo-EM to reach near-atomic resolution (3--5 Å) from small particles with low or even no symmetry. However, tools to determine structures from cryo-EM maps have lagged behind. In this dissertation we describe approaches that apply Rosetta high-resolution structure prediction methods to model-building from cryo-EM maps. Using knowledge-based information gathered from proteins of known atomic structure, we describe the development of novel tools for de novo model-building from near-atomic resolution cryo-EM maps, and for architecture determination of multi-component macromolecular assemblies from medium- to low-resolution (5--15 Å) cryo- EM maps. We demonstrate that these new computational tools have shown usefulness in determining the structures from newly reconstructed cryo-EM maps, which would otherwise be difficult or impossible for human to do. These methods should enable rapid and reliable structure determination from cryo-EM maps.

Electron Microscopy of Macromolecular Complexes, and Their Interactions with Membranes

Author : Rebecca Faith Thompson
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (118 download)

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Book Synopsis Electron Microscopy of Macromolecular Complexes, and Their Interactions with Membranes by : Rebecca Faith Thompson

Download or read book Electron Microscopy of Macromolecular Complexes, and Their Interactions with Membranes written by Rebecca Faith Thompson and published by . This book was released on 2015 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Scanning Electron Microscopy for the Life Sciences

Author : Heide Schatten
Publisher : Cambridge University Press
ISBN 13 : 0521195993
Total Pages : 275 pages
Book Rating : 4.5/5 (211 download)

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Book Synopsis Scanning Electron Microscopy for the Life Sciences by : Heide Schatten

Download or read book Scanning Electron Microscopy for the Life Sciences written by Heide Schatten and published by Cambridge University Press. This book was released on 2013 with total page 275 pages. Available in PDF, EPUB and Kindle. Book excerpt: A guide to modern scanning electron microscopy instrumentation, methodology and techniques, highlighting novel applications to cell and molecular biology.

Crystallization of Membrane Proteins

Author : Hartmut Michel
Publisher : CRC Press
ISBN 13 : 1351088173
Total Pages : 343 pages
Book Rating : 4.3/5 (51 download)

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Book Synopsis Crystallization of Membrane Proteins by : Hartmut Michel

Download or read book Crystallization of Membrane Proteins written by Hartmut Michel and published by CRC Press. This book was released on 2018-01-18 with total page 343 pages. Available in PDF, EPUB and Kindle. Book excerpt: The precise knowledge of the structure of biological macromolecules forms the basis of understanding their function and their mechanism of action. It also lays the foundation for rational protein and drug design. The only method to obtain this knowledge is still crystallography. At present, the structures of about 400 proteins are known at or nearly at atomic proteins. However, only two of them are membrane proteins or complexes of the membrane proteins. The reasons for the difference is not the crystals of membrane proteins resists forming special problems when being analysed. The reason is that the membrane proteins resist into forming into well-ordered crystals. The intention of this book is to help to produce well-ordered crystals proteins and to provide guidelines, it is aimed at both biochemists and protein crystallographer‘s.