Mutational Analysis of Gamma-tubulin PDF Download

Author: Robert Leo Jeng
Publisher:
ISBN:
Category :
Languages : en
Pages : 328

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Author: Jason Harris
Publisher:
ISBN:
Category :
Languages : en
Pages :

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"Inaccurate segregation of chromosomes during cell division is thought to lead to tumour formation. Microtubules play a key role in chromosome separation and segregation. The study of yeast spindles offers a convenient and versatile system to study the processes that impact cell division. One protein found to have an integral role in proper spindle assembly is yeast [gamma]-Tubulin (Tub4). Tub4 is a 53kDa protein that contains a flexible C-terminal tail. Removal of this tail has shown to be lethal in vivo. Mutation of a DYSL motif in the tail has also been shown to influence recruitment of proteins to the opposite end of a microtubule. Phosphomimetic mutations of the tyrosine in the DYSL motif have been shown to increase the number of microtubules that do not attach to kinetochores. NMR is ideally suited to study the effects of phosphorylation on the dynamics of the last 39 residues of Tub4. In doing so, we will gain information on how changes in the dynamics of Tub4 correlate to changes in protein function, which in turn influence the cell as a whole.We have used NMR spectroscopy to characterize the differences between the wild type(WT) Tub4 tail and a mutant containing the Y445D phosphomimetic substitution. Gradient diffusion experiments were employed to measure the hydrodynamic radii of the proteins, revealing that both WT and Y445D constructs diffuse more slowly than expected. The dynamics of the two constructs were analyzed using 15N and 2H spin relaxation measurements. Spectral density mapping and analysis of backbone dynamics show no change in the fast timescale dynamics by the Y445D mutation. Interestingly, CPMG analysis revealed that the Y445D mutation changed the milli-microsecond timescale dynamics of the protein. This change is observed throughout the entire protein and was determined to be a global exchange between a major and minor state. This suggests that the mutation causes the protein to exchange between collapsed and extended conformations. In this model, the extended conformation increases the capture radius in which it can interact with potential binding partners." --

Author: Anne Straube
Publisher: Humana Press
ISBN: 9781493961856
Category : Science
Languages : en
Pages : 319

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Microtubules are at the heart of cellular self-organization, and their dynamic nature allows them to explore the intracellular space and mediate the transport of cargoes from the nucleus to the outer edges of the cell and back. In Microtubule Dynamics: Methods and Protocols, experts in the field provide an up-to-date collection of methods and approaches that are used to investigate microtubule dynamics in vitro and in cells. Beginning with the question of how to analyze microtubule dynamics, the volume continues with detailed descriptions of how to isolate tubulin from different sources and with different posttranslational modifications, methods used to study microtubule dynamics and microtubule interactions in vitro, techniques to investigate the ultrastructure of microtubules and associated proteins, assays to study microtubule nucleation, turnover, and force production in cells, as well as approaches to isolate novel microtubule-associated proteins and their interacting proteins. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Definitive and practical, Microtubule Dynamics: Methods and Protocols provides the key protocols needed by novices and experts on how to perform a broad range of well-established and newly-emerging techniques in this vital field.

Author:
Publisher: Elsevier
ISBN: 0323140068
Category : Nature
Languages : en
Pages : 482

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This highly researched yeast, which represents a system used by cell biologists, geneticists and molecular biologists, has been given only minimal coverage in the literature. Its properties make it an excellent organism for DNA and related biotechnology reseach. This book, which is the first attempt to collate existing information in one source, will be an invaluable aid to those initiating projects with this organism.

Author:
Publisher:
ISBN: 9780815332183
Category : Cells
Languages : en
Pages : 0

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Author: Mohan L. Gupta
Publisher:
ISBN:
Category :
Languages : en
Pages : 358

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Author: Wei Yamauchi
Publisher: Nova Science Publishers
ISBN: 9781621001416
Category : Tubulins
Languages : en
Pages : 0

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Tubulin is one of several members of a small family of globular proteins. The most common members of the tubulin family are α-tubulin and β-tubulin, the proteins that make up microtubules. In this book the authors present topical research in the study of the structure, functions and roles in disease of tubulin. Topics discussed in this compilation include microtubule plus-end tracking proteins in neuronal development and nervous system disorders; trypanosome tubulin molecules as targets for drug and vaccine development for protozoan infections; tubulin inhibitors as vascular disrupting agents and evolution and co-evolution of tubulin's carboxy-terminal tails and mitochondria.

Author:
Publisher: Academic Press
ISBN: 0123813506
Category : Science
Languages : en
Pages : 549

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Book Description
Microtubules: in vivo includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton. The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation. - Includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton - The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation

Author: Antonio Tito Fojo
Publisher: Springer Science & Business Media
ISBN: 1597453366
Category : Medical
Languages : en
Pages : 629

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Book Description
This book presents the first comprehensive exploration of the dynamic potential of microtubules anti-cancer targets. Written by leading anti-cancer researchers, this groundbreaking volume collects the most current microtubule research available and investigates the potential of microtubules in cancer therapy.

Author: John J. Correia
Publisher: Academic Press
ISBN: 0124078885
Category : Science
Languages : en
Pages : 472

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Book Description
There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that control cell cycle, dynamics, organization and transport. The field continues to grow and explore new aspects of these issues driven immensely by developments in optical imaging and tracking techniques. This Second Edition brings together current research and protocols in the field of microtubules in vitro and will serve as a valuable tool for cell biologists, biophysicists and pharmacologists who study the microtubule cytoskeleton, as well as for researchers in the biomedical and biotechnology communities with interest in developing drugs that target microtubules, MAPS and motors. - Chapters reflect experimental procedures and new developments in the field of microtubule in vitro research - Combines classical approaches and modern technologies - Presents easy-to-use protocols and thorough background information, compiled by leaders in the field