Download or read book Development of In situ Protein Engineering Technologies for the Study of Modified Proteins written by Michael R. Haugbro and published by . This book was released on 2021 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Deciphering the roles that protein post-translational modifications, protein-protein interactions, and protein dynamics play in modulating protein structure and function requires the ability to introduce amino acid modifications, cross-linking moieties, biophysical probes, and other synthetic chemical species into proteins. Current technologies for the semi-synthesis of modified proteins are optimized for in vitro reactions and are not easily transferable to more biologically relevant systems such as live cell culture. Analogous methodologies that work in situ rely on genetic manipulations that often result in low resolution data and a high level of off-target effects. Early attempts to address this disconnect have employed protein ligase systems such as Sortase and split inteins, but as of yet have not become robust enough for wide-spread use. In this thesis we expand upon the existing technologies and introduce new methods for the precise chemical manipulation of proteins in cell culture using split inteins. We applied in-nucleo protein trans-splicing to determine the in situ interactome of histone PTMs, combining chemical precision and native protein environment in an unprecedented manner unattainable by any other approach. The full results of the proteomics experiments enabled by this technique can be found in Appendix 2, available with the digital version of this thesis. We also expanded the in situ PTS toolbox by developing a multiplexed method for histone labeling in nuclei and in live cells. We demonstrated this capability by labeling two histones with different color fluorophores in a completely orthogonal manner. Finally, we utilized an atypical split intein known as VidaL to achieve the first semi-synthesis of a modified protein in cells, generating histones bearing multiple PTMs and an affinity handle in the chromatin of live cells. We added to this platform by combining our semi-synthesis capabilities with control of the subcellular localization of our target protein, in a single reaction. We envision that these new methods described in this thesis will equip protein chemists and cellular biologists alike with new capabilities for the study of protein structure and function.
Download or read book Protein Engineering For Industrial Biotechnology written by Lilia Alberghina and published by CRC Press. This book was released on 2003-09-02 with total page 374 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein engineering has proved to be one of the more fruitful technological approaches in biotechnology, being both very powerful and able to generate valuable intellectual property. This book aims to present examples in which the application of protein engineering has successfully solved problems arising in industrial biotechnology. There is a section on its use to enhance purification of recombinant proteins. The use of protein engineering to modify the activity or the stability of industrial enzymes from lipases to proteases, from carboxypeptidases to glucanases and glucosidases, and from pectin modifying enzymes to enzymes able to degrade recalcitrant compounds is extensively covered. It is shown how areas as diverse as agrofood technology, fine chemistry, detergents, bioremediation and biosensors receive significant contributions from protein and solvent engineering. The application of protein engineering to health care is also covered, from the development of new vaccines to new potential therapeutic proteins. A specific notation is given to protein engineering in the development of target molecules for drug discovery. International in scope, the many contributions are drawn from academia and industry. The text should be of interest to students and researchers in industrial biotechnology as well as to everybody interested in basic research in protein structure, molecular genetics, bio-organic chemistry, biochemistry, agrobiotechnology, pharmaceutical sciences and medicine.
Download or read book Protein Engineering written by Caroline Koehrer and published by Springer Science & Business Media. This book was released on 2009-01-07 with total page 351 pages. Available in PDF, EPUB and Kindle. Book excerpt: Site-specific mutagenesis of DNA, developed some thirty years ago, has proven to be one of the most important advances in biology. By allowing the site-specific replacement of any amino acid in a protein with one of the other nineteen amino acids, it ushered in the new era of "Protein Engineering". The field of protein engineering has, however, evolved rapidly since then and the last fifteen years have witnessed remarkable advances through the use of new chemical, biochemical and molecular biological tools towards the synthesis and manipulation of proteins. The chapters included in this book reflect the rapid evolution of protein engineering and its many applications in basic research, biotechnology, material sciences and therapy. This book will provide the reader with an introduction to state-of the-art concepts and methods and will be of use to anyone interested in the study of proteins, in academia as well as in industry.
Download or read book Protein Engineering Protocols written by Kristian Müller and published by Springer Science & Business Media. This book was released on 2007-10-26 with total page 318 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein engineering is a fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry, with the goal of developing useful or valuable proteins. Protein Engineering Protocols will consider the two general, but not mutually exclusive, strategies for protein engineering. The first is known as rational design, in which the scientist uses detailed knowledge of the structure and function of the protein to make desired changes. The s- ond strategy is known as directed evolution. In this case, random mutagenesis is applied to a protein, and selection or screening is used to pick out variants that have the desired qualities. By several rounds of mutation and selection, this method mimics natural evolution. An additional technique known as DNA shuffling mixes and matches pieces of successful variants to produce better results. This process mimics recombination that occurs naturally during sexual reproduction. The first section of Protein Engineering Protocols describes rational p- tein design strategies, including computational methods, the use of non-natural amino acids to expand the biological alphabet, as well as impressive examples for the generation of proteins with novel characteristics. Although procedures for the introduction of mutations have become routine, predicting and und- standing the effects of these mutations can be very challenging and requires profound knowledge of the system as well as protein structures in general.
Download or read book Protein Engineering written by Tomohisa Ogawa and published by BoD – Books on Demand. This book was released on 2013-05-29 with total page 198 pages. Available in PDF, EPUB and Kindle. Book excerpt: Given the centrality of protein to many biological process, this book makes a significant contribution to the fields of healthcare and nutrition. Its chapters consider topics such as protein-protein and protein-ligand docking, and the protein engineering of enzymes involved in bioplastic metabolism. One contribution gives an overview of the In Vitro Virus (IVV) analytic method, while another shows how cutting-edge techniques in protein engineering advance our knowledge in the field of palaeontology. The book also includes a review of classic and alternative strategies when using yeasts in research, with a focus on Pichia pastoris as a host. Finally, there are two contributions on chromatography: one on the method itself, and another on its use to identify HMGB1-binding components.
Download or read book Protein Engineering by Semisynthesis written by Carmichael J.A. Wallace and published by CRC Press. This book was released on 1999-09-29 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: As genomics gives way to proteomics as the focus of scientific imagination in the biological sciences, more emphasis will be placed on the technology and interpretation of protein engineering experiments. Protein engineers will become increasingly sophisticated in the questions that they pose and demanding of the tools available to change protein structure. The optimal way to introduce non-coded amino acids for mechanistic studies, or site-specific reporter atoms for spectroscopic structural biology, is by protein semisynthesis. In Protein Engineering by Semisynthesis, the leading practitioners of the method cover their individual protein of expertise forming a comprehensive illustration of the various methods developed. By covering the most recent philosophical and methodological approaches and developments of semisynthesis and peptide synthesis to date, this book provides further understanding of the principles of protein structure-function relationships gained from semisynthetic analog in addition to providing a comprehensive and comprehensible laboratory guide. This book focuses on recent developments which synergistically combine chemical and molecular biological techniques that have made semisynthetic manipulations much easier to undertake. Features
Download or read book Protein Engineering written by Jeffrey L. Cleland and published by Wiley-Liss. This book was released on 1996-02-07 with total page 558 pages. Available in PDF, EPUB and Kindle. Book excerpt: This systematic approach to protein engineering provides a thorough introduction to protein biophysics via chapters dealing with the experimental and theoretical determination of protein structure. Separate chapters discuss each of the expression systems c
Download or read book Protein Engineering Techniques written by Krishna Mohan Poluri and published by Springer. This book was released on 2016-11-12 with total page 144 pages. Available in PDF, EPUB and Kindle. Book excerpt: This brief provides a broad overview of protein-engineering research, offering a glimpse of the most common experimental methods. It also presents various computational programs with applications that are widely used in directed evolution, computational and de novo protein design. Further, it sheds light on the advantages and pitfalls of existing methodologies and future perspectives of protein engineering techniques.
Download or read book Protein Engineering written by Caroline Koehrer and published by Springer. This book was released on 2009-08-29 with total page 347 pages. Available in PDF, EPUB and Kindle. Book excerpt: Site-specific mutagenesis of DNA, developed some thirty years ago, has proven to be one of the most important advances in biology. By allowing the site-specific replacement of any amino acid in a protein with one of the other nineteen amino acids, it ushered in the new era of "Protein Engineering". The field of protein engineering has, however, evolved rapidly since then and the last fifteen years have witnessed remarkable advances through the use of new chemical, biochemical and molecular biological tools towards the synthesis and manipulation of proteins. The chapters included in this book reflect the rapid evolution of protein engineering and its many applications in basic research, biotechnology, material sciences and therapy. This book will provide the reader with an introduction to state-of the-art concepts and methods and will be of use to anyone interested in the study of proteins, in academia as well as in industry.
Download or read book Protein Engineering written by Dan Robertson and published by Elsevier. This book was released on 2004-09-29 with total page 453 pages. Available in PDF, EPUB and Kindle. Book excerpt: This MIE volume covers methods for a multitude of topics among which are computational methods, laboratory methods, enzyme optimization, binding proteins/antibodies, and screening technologies. Table of Contents-Methodology-Applications-Opzimization and Screening-Applications-Directed Evolution of Enzymatic Function-Applications-Evolution of Biosynthetic Pathways-Devices, Antibodies and Vaccines
Download or read book Protein Engineering written by Raghupathy Sarma and published by Elsevier. This book was released on 2012-12-02 with total page 441 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein Engineering: Applications in Science, Medicine, and Industry deals with the scientific, medical, and industrial applications of protein engineering. Topics range from protein structure and design to mutant analysis and complex systems. Applications such as production of novel antibiotics, genetic transformation of plants, and genetic engineering of bioinsecticides are described. This book is comprised of 25 chapters and begins with an overview of trends and developments in protein chemistry and their relevance to protein engineering, followed by a discussion on protein sequence data banks. Subsequent chapters explore the design and construction of biologically active peptides, including hormones; structural and functional analysis of thermophile proteins; the conformation of diphtheria toxin; and applications of surface-simulation synthesis in protein molecular recognition. The use of oligonucleotide-directed site-specific mutagenesis in functional analysis of the signal peptide for protein secretion is also considered. The results of studies on the mechanism of membrane fusion are presented. This monograph will serve as a useful guide for those who are already working on protein engineering and those who are about to start research in this field.
Download or read book Protein Engineering and Design written by Paul R. Carey and published by Elsevier. This book was released on 1996-06-21 with total page 379 pages. Available in PDF, EPUB and Kindle. Book excerpt: The design and production of novel peptides and proteins occupy pivotal positions in science and technology and will continue to do so in the 21st century. Protein Engineering and Design outlines the rapid advances in computer-based modeling, protein engineering, and methods needed for protein and peptide preparation and characterization. This indispensable reference lays the groundwork for understanding this multidisciplinary activity while providing an introduction for researchers and students to the field of protein design. Introduces and defines the techniques involved in protein engineering and design Provides a concise overview of key technologies involved and demonstrates their contributions to the specialized design and production of novel proteins and peptides
Download or read book Protein Engineering written by Huimin Zhao and published by John Wiley & Sons. This book was released on 2021-08-23 with total page 41 pages. Available in PDF, EPUB and Kindle. Book excerpt: A one-stop reference that reviews protein design strategies to applications in industrial and medical biotechnology Protein Engineering: Tools and Applications is a comprehensive resource that offers a systematic and comprehensive review of the most recent advances in the field, and contains detailed information on the methodologies and strategies behind these approaches. The authors—noted experts on the topic—explore the distinctive advantages and disadvantages of the presented methodologies and strategies in a targeted and focused manner that allows for the adaptation and implementation of the strategies for new applications. The book contains information on the directed evolution, rational design, and semi-rational design of proteins and offers a review of the most recent applications in industrial and medical biotechnology. This important book: Covers technologies and methodologies used in protein engineering Includes the strategies behind the approaches, designed to help with the adaptation and implementation of these strategies for new applications Offers a comprehensive and thorough treatment of protein engineering from primary strategies to applications in industrial and medical biotechnology Presents cutting edge advances in the continuously evolving field of protein engineering Written for students and professionals of bioengineering, biotechnology, biochemistry, Protein Engineering: Tools and Applications offers an essential resource to the design strategies in protein engineering and reviews recent applications.
Download or read book Biobetters written by Amy Rosenberg and published by Springer. This book was released on 2015-08-23 with total page 378 pages. Available in PDF, EPUB and Kindle. Book excerpt: “Biobetters: Protein Engineering to Approach the Curative” discusses the optimization of protein therapeutic products for treatment of human diseases. It is based on the fact that though numerous important therapeutic protein products have been developed for life threatening and chronic diseases that possess acceptable safety and efficacy profiles, these products have generally not been reexamined and modified for an improved clinical performance, with enhancements both to safety and efficacy profiles. Advances in protein engineering, coupled with greatly enhanced understanding of critical product quality attributes for efficacy and safety, make it possible to optimize predecessor products for clinical performance, thereby enhancing patient quality of life and with the potential for great savings in health care costs. Yet despite such knowledge, there is little movement towards such modifications. This book examines engineering protein therapeutic products such that they exhibit an optimal, not just an adequate, clinical performance profile. Two product classes, therapeutic enzymes for lysosomal storage diseases (enzyme replacement therapies, ERT) and monoclonal antibodies (mAbs), are used as examples of what modifications to such proteins could be made to enhance clinical performance, “closer to a cure” as it were. For ERT, the key to optimizing clinical performance is to ensure the ERT is endowed with moieties that target the protein to the relevant target tissue. Thus, for Gaucher Disease, our best example of how to optimize an ERT to address a disease that manifests in specific target tissues (macrophages and monocytes), the enzyme has been extensively modified to target macrophages. For diseases such as Pompe Disease, largely a disorder of muscle, optimal performance of ERT will depend on endowing the enzyme with the ability to be taken up via the Mannose 6 Phosphate Receptor, and so one of the chapters in the book will discuss such approaches. Moreover, a major failure of biotechnology based products is to gain access to the CNS, a key target tissue in numerous diseases. Thus, a chapter has been devoted to strategies to access the CNS. Additionally, immune responses to therapeutic proteins can be highly problematic, eliminating the efficacy of life saving or highly effective protein therapeutics. This is especially poignant in the case of Pompe Disease wherein great improvement in muscle strength and functionality is lost following development of an immune response to the ERT with consequent patient deterioration and death. Thus, a chapter regarding protein engineering, as well as other non-clinical approaches to diminishing immunogenicity is a valuable part of the book. Monoclonal antibodies (mAbs) can be engineered to bind targets relevant to a wide variety of diseases; binding affinity, however, is only part of the equation and one of the chapters will present a molecular assessment approach that balances affinity with pharmacokinetics and manufacturability. As with other proteins immunogenicity can be problematic, being responsible for loss of efficacy of anti-TNF mAbs, often after prolonged successful treatment. The authors will also share their perspective on the consequences of physico-chemical modifications occurring to mAbs once they reach the circulation or their target, a research area open to further development from a protein engineering as well as analytical perspective. This book will also discuss novel platforms for protein therapeutics, technologies that exceed mAbs with respect to potency, and hence, potentially efficacy. These platforms consist largely of repeat domain proteins with very high affinity for their target ligands, but while potentially more efficacious, immunogenicity may be a major problem limiting use. The economics surrounding the issue of biobetters is another high-profile issue - this final chapter will explore the incentives and disincentives for developing biobetters and consider incentives that might make their pursuit more rewarding.
Download or read book Concepts in Protein Engineering and Design written by Paul Wrede and published by Walter de Gruyter GmbH & Co KG. This book was released on 2020-10-12 with total page 396 pages. Available in PDF, EPUB and Kindle. Book excerpt: No detailed description available for "Concepts in Protein Engineering and Design".
Download or read book A Practical Guide to Protein Engineering written by Tuck Seng Wong and published by Springer Nature. This book was released on 2020-10-29 with total page 213 pages. Available in PDF, EPUB and Kindle. Book excerpt: This textbook introduces readers in an accessible and engaging way to the nuts and bolts of protein expression and engineering. Various case studies illustrate each step from the early sequence searches in online databases over plasmid design and molecular cloning techniques to protein purification and characterization. Furthermore, readers are provided with practical tips to successfully pursue a career as a protein engineer. With protein engineering being a fundamental technique in almost all molecular biology labs, the book targets advanced undergraduates and graduate students working in molecular biology, biotechnology and related scientific fields.
Download or read book Methods in Protein Design written by and published by Academic Press. This book was released on 2013-02-14 with total page 539 pages. Available in PDF, EPUB and Kindle. Book excerpt: This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. This volume covers methods in protein design and it has chapters on such topics as protein switch engineering by domain insertion, evolution based design of proteins, and computationally designed proteins. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers methods in protein design Contains chapters with such topics as protein switch engineering by domain insertion, evolution-based design of proteins, and computationally designed proteins
