Isolation and Structural Characterization of a Subset of Yeast (Saccharomyces Cerevisiae) Peroxisomal Proteins PDF Download
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Author: Munmun S. Nandi
Publisher:
ISBN:
Category :
Languages : en
Pages : 202
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Book Description
Peroxisomes are virtually found in all eukaryotic cells, but unlike mitochondria and chloroplasts, they do not contain DNA or a protein secretory apparatus. Therefore, all of their proteins must be imported by a process called peroxisomal biogenesis. This requires a group of protein factors referred to as peroxins which are encoded by the pex genes. Currently, there are approximately thirty-two known peroxisomal proteins. Among all the peroxisomal proteins, two enzymes namely GPD1, LYS1 and a peroxin, PEX7 were selected for the research. GPD1 is a NAD+ -dependent glycerol 3-phosphate dehydrogenase1 that catalyzes the conversion of dihydroxyacetone phosphate (DHAP) to glycerol 3-phosphate which is crucial for growth under osmotic stress. Its purification was achieved using ion exchange chromatography and the pure protein was crystallized for structure determination. Diffraction data sets were obtained to a resolution of 2.2 Å which were used to solve the C-terminal portion of the structure. Unfortunately, the N-terminal portion remained disordered. LYS1 is the terminal enzyme of [alpha]-aminoadipate pathway and catalyzes the reversible NAD-dependent oxidative cleavage of saccharopine to yield L-lysine and [alpha]-ketoglutarate. The purification of LYS1 was carried out using affinity chromatography. Another protein, PEX7 is responsible for peroxisome biogenesis by importing newly synthesized proteins bearing PTS2 (peroxisome targeting signal sequence2) into peroxisomes. PEX7 presented the greatest challenge among the three proteins at both the expression stage and the purification stage. Its soluble fraction was purified using ion exchange and affinity chromatographies, although the final yield was too low for crystallization trials. A much large proportion of the protein was found in the insoluble cell debris and attempts were made to purify this fraction after denaturation. An alternative, protocol involving the formation of a GPD1-PEX7 complex proved to be effective route to co-purification of the two proteins and crystallization trials are proceeding. Having known the structures of peroxisomal proteins, it would be helpful for studying the development and maintenance of the organelle related to its metabolic diseases in the eukaryotic cells.
Author: Munmun S. Nandi
Publisher:
ISBN:
Category :
Languages : en
Pages : 202
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Book Description
Peroxisomes are virtually found in all eukaryotic cells, but unlike mitochondria and chloroplasts, they do not contain DNA or a protein secretory apparatus. Therefore, all of their proteins must be imported by a process called peroxisomal biogenesis. This requires a group of protein factors referred to as peroxins which are encoded by the pex genes. Currently, there are approximately thirty-two known peroxisomal proteins. Among all the peroxisomal proteins, two enzymes namely GPD1, LYS1 and a peroxin, PEX7 were selected for the research. GPD1 is a NAD+ -dependent glycerol 3-phosphate dehydrogenase1 that catalyzes the conversion of dihydroxyacetone phosphate (DHAP) to glycerol 3-phosphate which is crucial for growth under osmotic stress. Its purification was achieved using ion exchange chromatography and the pure protein was crystallized for structure determination. Diffraction data sets were obtained to a resolution of 2.2 Å which were used to solve the C-terminal portion of the structure. Unfortunately, the N-terminal portion remained disordered. LYS1 is the terminal enzyme of [alpha]-aminoadipate pathway and catalyzes the reversible NAD-dependent oxidative cleavage of saccharopine to yield L-lysine and [alpha]-ketoglutarate. The purification of LYS1 was carried out using affinity chromatography. Another protein, PEX7 is responsible for peroxisome biogenesis by importing newly synthesized proteins bearing PTS2 (peroxisome targeting signal sequence2) into peroxisomes. PEX7 presented the greatest challenge among the three proteins at both the expression stage and the purification stage. Its soluble fraction was purified using ion exchange and affinity chromatographies, although the final yield was too low for crystallization trials. A much large proportion of the protein was found in the insoluble cell debris and attempts were made to purify this fraction after denaturation. An alternative, protocol involving the formation of a GPD1-PEX7 complex proved to be effective route to co-purification of the two proteins and crystallization trials are proceeding. Having known the structures of peroxisomal proteins, it would be helpful for studying the development and maintenance of the organelle related to its metabolic diseases in the eukaryotic cells.
Author: Cecile Brocard
Publisher: Springer
ISBN: 3709117887
Category : Science
Languages : en
Pages : 540
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Book Description
In eukaryotes, lipid metabolism requires the function of peroxisomes. These multitasking organelles are also part of species-specific pathways such as the glyoxylate cycle in yeast and plants or the synthesis of ether lipid in mammals. Proteins required for the biogenesis of peroxisomes typically assemble in large molecular complexes, which participate in membrane formation, protein transport, peroxisome duplication and - inheritance during cell division. Peroxisomal function is essential for life. Mutations in PEX genes, encoding for biogenesis factors, are often associated with lethal disorders. The association of peroxisomes with other organelles suggests an extensive participation in organellar crosstalk. This book represents a state-of-the-art review in the field of peroxisome research encompassing the cell and molecular biology of peroxisome biogenesis and its diseases, the protein complexes involved in this process and the modern technologies applied to study them. The book is intended for graduate students, researchers and lecturers in biochemistry, molecular and cell biology with a biomedical background.
Author: L.A. Grivell
Publisher: Springer Science & Business Media
ISBN: 940112504X
Category : Science
Languages : en
Pages : 154
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Book Description
At a fundamental research level, the yeasts offer valuable opportunities for modelling regulatory and metabolic processes in multicellular eukaryotic organisms: this volume deals with the multifunctional chromosome regulatory proteins, topoisomerase and nuclear transport. A combination of biochemical and genetic approaches applied to the yeast translation system is also currently yielding a wealth of data, while the mating pheromone signal transduction pathway in yeasts provides a valuable analogue of the signal transduction components used by multicellular organisms, including receptors, G proteins, protein kinases and transcription factors. With a well-established history of fermantation studies, yeasts remain the first-choice vehicle for production of heterologous eukaryotic proteins. Interest is diversifying, as an increasing number of non-Saccharomyces species are now being utilised for the production of specific heterologous proteins. Molecular biologists, microbiologists and biochemical geneticists will find this volume an authoritative and valuable update on a vibrant area of research.
Author: Luis A. del Río
Publisher: Springer Science & Business Media
ISBN: 9400768893
Category : Medical
Languages : en
Pages : 353
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Book Description
Peroxisomes are a class of ubiquitous and dynamic single membrane-bounded cell organelles, devoid of DNA, with an essentially oxidative type of metabolism. In recent years it has become increasingly clear that peroxisomes are involved in a range of important cellular functions in almost all eukaryotic cells. In higher eukaryotes, including humans, peroxisomes catalyze ether phospholipids biosynthesis, fatty acid alpha-oxidation, glyoxylate detoxification, etc, and in humans peroxisomes are associated with several important genetic diseases. In plants, peroxisomes carry out the fatty acid beta-oxidation, photorespiration, metabolism of ROS, RNS and RSS, photomorphogenesis, biosynthesis of phytohormones, senescence, and defence against pathogens and herbivores. In recent years it has been postulated a possible contribution of peroxisomes to cellular signaling. In this volume an updated view of the capacity and function of peroxisomes from human, animal, fungal and plant origin as cell generators of different signal molecules involved in distinct processes of high physiological importance is presented.
Author: Vladimir I Titorenko
Publisher: Frontiers Media SA
ISBN: 2889194647
Category : Physiology
Languages : en
Pages : 149
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Book Description
The peroxisome is an organelle with essential roles in lipid metabolism, maintenance of reactive oxygen species homeostasis, and anaplerotic replenishment of tricarboxylic acid cycle intermediates destined for mitochondria. Peroxisomes constitute a dynamic endomembrane system. The homeostatic state of this system is upheld via two pathways for assembling and maintaining the diverse peroxisomal compartments constituting it; the relative contribution of each pathway to preserving such system may vary in different organisms and under various physiological conditions. One pathway begins with the targeting of certain peroxisomal membrane proteins to an endoplasmic reticulum template and their exit from the template via pre-peroxisomal carriers; these carriers mature into metabolically active peroxisomes containing the entire complement of membrane and matrix proteins. Another pathway operates via growth and maturation of pre-existing peroxisomal precursors that do not originate from the endoplasmic reticulum; mature peroxisomes proliferate by undergoing fission. Recent studies have uncovered new roles for the peroxisomal endomembrane system in orchestrating important developmental decisions and defining organismal longevity. This Frontiers Special Topic Issue is focused on the advances in our understanding of how evolutionarily distant organisms coordinate the formation, maturation, proliferation, maintenance, inheritance and quality control of the peroxisomal endomembrane system and how peroxisomal endomembranes communicate with other cellular compartments to orchestrate complex biological processes and various developmental programs from inside the cell.
Author: Allan Peter Spong
Publisher:
ISBN:
Category :
Languages : en
Pages : 308
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Book Description
Author: A. Baker
Publisher: Springer
ISBN: 9789401598590
Category : Science
Languages : en
Pages : 505
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Book Description
In the two decades since the last comprehensive work on plant peroxisomes appeared, the scientific approaches employed in the study of plant biology have changed beyond all recognition. The accelerating pace of plant research in the post-genomic era is leading us to appreciate that peroxisomes have many important roles in plant cells, including reserve mobilisation, nitrogen assimilation, defence against stress, and metabolism of plant hormones, which are vital for productivity and normal plant development. Many plant scientists are finding, and will no doubt continue to find, that their own area of research is connected in some way to peroxisomes. Written by the leading experts in the field, this book surveys peroxisomal metabolic pathways, protein targeting and biogenesis of the organelle and prospects for the manipulation of peroxisomal function for biotechnological purposes. It aims to draw together the current state of the art as a convenient starting point for anyone, student or researcher, who wishes to know about plant peroxisomes.
Author: Tsuneo Imanaka
Publisher: Springer Nature
ISBN: 9811511691
Category : Medical
Languages : en
Pages : 284
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Book Description
This book provides readers with a comprehensive overview of peroxisomes and their role in human diseases. It starts by describing the history of peroxisome research and then examines in detail the current understanding of the biogenesis and function of peroxisomes. It then focuses on peroxisomal disorders and the involvement of peroxisomes in cancer and age-related diseases, discussing in detail the use of model organisms to elucidate the pathogenesis of peroxisomal disorders and the physiological importance of peroxisomal proteins. Further, the book examines diagnostic and therapeutic strategies in peroxisomal disorders as well as significant recent advances. Lastly, it addresses various topics in peroxisome research, including the isolation of peroxisomes from mammalian tissues and cells, the structural biology of peroxisomal proteins, the lipidomics of peroxisomal disorders, the value of exome sequencing, and neuropsychological testing in X-linked adrenoleukodystrophy. Given its scope, the book is a valuable resource for postgraduate students and researchers in the life sciences and clinicians in the fields of internal medicine, pediatrics, and neurology.
Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1844
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Book Description
Author: Waleed Mohamed Hussain Abdulkhair
Publisher: BoD – Books on Demand
ISBN: 9535137344
Category : Technology & Engineering
Languages : en
Pages : 178
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Book Description
Biotechnology including medical applications depends on the yeast as biofermenter to produce many industrial products including pharmaceutical ones. Although yeasts are first known as useful microorganisms, some of them are identified as pathogens for plants, animals, and humans. Due to the simple cellular structure of the yeast among other microbial groups, it is used in the earliest investigations to determine the features of eukaryotic molecular biology, cell biology, and physiology. The economic income of some countries mainly depends on yeast for producing the economic products, such as France that depends on yeast for wine production. This book throws light on yeast and its important role in the medical applications.
