Download or read book Hybrid phase Native Chemical Ligation Approaches to Overcome the Limitations of Protein Total Synthesis written by Ruixuan Ryan Yu and published by . This book was released on 2016 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Total protein synthesis allows the preparation of proteins with chemically diverse modifications. The numerous advantages of total synthesis are sometimes offset by some major limitations. Protein synthesis is a non-trivial task involving many chemical steps, and these steps increase with the size of the protein. Therefore, larger proteins are difficult to synthesize with high yield. We have developed a strategy which we term hybrid-phase native chemical ligation (NCL) to overcome some of the limitations of size and yield. Hybrid-phase NCL combines ligating peptides on a solid support (solid-phase NCL) and in solution (solution-phase NCL) to maximize synthetic yield. We have successfully used this method to synthesize triple-acetylated histone H4-K5ac,K12ac,K91ac and, for the first time, acetylated centromeric histone CENP-A-K124ac (CpA-K124ac). In order to improve the yield of CENP-A total synthesis, we have incorporated a convergent ligation element in our hybrid-phase strategy. This new approach reduced the number of purification steps, leading to a synthetic yield that was almost triple that of the original approach. Finally, we introduce the convergent solid-phase hybrid NCL approach that allows the preparation of a long peptide segment bearing a masked thioester on a solid support. Through a newly developed resin-anchoring strategy, cleavage of the product from solid-phase generated a ligation-compatible segment that could be used directly with no purification. This method has the potential to synthesize large proteins in good yield, effectively overcoming the size and yield limits of protein total synthesis.
Download or read book Convergent Hybrid phase Ligation Synthetic Strategy Combined with Direct Refolding Method to Improve Synthetic Yield of Centromeric Protein A written by Xiaoyu Zhang and published by . This book was released on 2020 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Histone proteins are a series of post-translational modification (PTM) heavy proteins. Total protein chemical synthesis possesses advantages in preparation of these protein samples containing specific PTM(s). However, the synthetic yield is always one of the biggest limitation of total protein chemical synthesis, especially when preparing large protein sample. Our lab had developed a synthetic strategy to total synthesize the H3 variant, Centromeric protein A (Cenp- A). Even though the synthesis was successful, the overall synthetic yield still did not fulfil our expectation. Here, we have developed a new convergent hybrid-phase synthetic strategy combined with direct histone octamer refolding and applied on the total synthesis of Cenp-A. This new strategy could maximally utilize the advantage of solid-phase native chemical ligation and avoid the any solution-phase purification steps which turned out to be the main reason of causing yield loss during the synthesis. This new synthetic strategy holds potential of greatly improving the synthetic yield of Cenp-A and being applied on the total synthesis of other large proteins.
Download or read book Chemical Ligation written by Luca D. D'Andrea and published by John Wiley & Sons. This book was released on 2017-03-27 with total page 558 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presenting a wide array of information on chemical ligation – one of the more powerful tools for protein and peptide synthesis – this book helps readers understand key methodologies and applications that protein therapeutic synthesis, drug discovery, and molecular imaging. • Moves from fundamental to applied aspects, so that novice readers can follow the entire book and apply these reactions in the lab • Presents a wide array of information on chemical ligation reactions, otherwise scattered across the literature, into one source • Features comprehensive and multidisciplinary coverage that goes from basics to advanced topics • Helps researchers choose the right chemical ligation technique for their needs
Download or read book Development of Convergent Hybrid Phase Ligation for Chemical Synthesis of Challenging Proteins written by Ziyong Hong (Ph. D. in chemistry) and published by . This book was released on 2021 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Chemical protein synthesis has proven to be an important tool for investigation of protein structure and function, but application to large proteins remains a major challenge. Here we develop convergent hybrid phase native chemical ligation (CHP-NCL) as a new method for practical preparation of large proteins from multiple small, synthetically accessible peptide segments. This strategy combines the best features of ligation on the solid phase, to obtain reactive peptide “blocks” approximately 100 residues in length from several short peptide segments, and concepts from convergent ligation, which bring these blocks together for further ligation steps in solution phase. The parallel preparation of each block greatly improves the efficiency and overall yield of protein synthesis. We further carefully investigate the possible side reactions observed in the use of various derivatives of 3,4-aminobenzoic acid (Dbz), an enabling chemical functional group for this strategy, and provide guidance for the selection and use of appropriate variants in the preparation of activated peptide N-acylureas and thioesters. We demonstrate the feasibility of our CHP-NCL strategy on the synthesis of the 212-residue linker histone H1.2, with several combinations of post-translational modifications (PTMs) including citrullination, phosphorylation, and acetylation at sites that are hypothesized to play important roles in the regulation of chromatin structure and dynamics. We provide preliminary biochemical and biophysical characterization of the effect of these modifications on the interaction of H1.2 with nucleosomes. We anticipate that this new approach will find wide applications in the preparation of large proteins and facilitate the understanding of protein structure and function.
Download or read book Chemical Approaches to the Synthesis of Peptides and Proteins written by Paul Lloyd-Williams and published by CRC Press. This book was released on 1997-04-23 with total page 306 pages. Available in PDF, EPUB and Kindle. Book excerpt: Organic chemists working on the synthesis of natural products have long found a special challenge in the preparation of peptides and proteins. However, more reliable, more efficient synthetic preparation methods have been developed in recent years. This reference evaluates the most important synthesis methods available today, and also considers methods that show promise for future applications. This text describes the state of the art in efficient synthetic methods for the synthesis of both natural and artificial large peptide and protein molecules. Subjects include an introduction to basic topics, linear solid-phase synthesis of peptides, peptide synthesis in solution, convergent solid-phase synthesis, methods for the synthesis of branched peptides, formation of disulfide bridges, and more. The book emphasizes strategies and tactics that must be considered for the successful synthesis of peptides.
Download or read book Total Chemical Synthesis of Proteins written by Ashraf Brik and published by John Wiley & Sons. This book was released on 2021-06-08 with total page 626 pages. Available in PDF, EPUB and Kindle. Book excerpt: How to synthesize native and modified proteins in the test tube With contributions from a panel of experts representing a range of disciplines, Total Chemical Synthesis of Proteins presents a carefully curated collection of synthetic approaches and strategies for the total synthesis of native and modified proteins. Comprehensive in scope, this important reference explores the three main chemoselective ligation methods for assembling unprotected peptide segments, including native chemical ligation (NCL). It includes information on synthetic strategies for the complex polypeptides that constitute glycoproteins, sulfoproteins, and membrane proteins, as well as their characterization. In addition, important areas of application for total protein synthesis are detailed, such as protein crystallography, protein engineering, and biomedical research. The authors also discuss the synthetic challenges that remain to be addressed. This unmatched resource: Contains valuable insights from the pioneers in the field of chemical protein synthesis Presents proven synthetic approaches for a range of protein families Explores key applications of precisely controlled protein synthesis, including novel diagnostics and therapeutics Written for organic chemists, biochemists, biotechnologists, and molecular biologists, Total Chemical Synthesis of Proteins provides key knowledge for everyone venturing into the burgeoning field of protein design and synthetic biology.
Download or read book Solid Phase Peptide Synthesis written by Gregg B. Fields and published by Academic Press. This book was released on 1997-10-21 with total page 828 pages. Available in PDF, EPUB and Kindle. Book excerpt: The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volumehas been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. More than 275 volumes have been published (all of them still in print) and much of the material is relevant even today-truly an essential publication for researchers in all fields of life sciences. Key Features * Solid-phase peptide synthesis * Applications of peptides for structural and biological studies * Characterization of synthetic peptides
Download or read book Plant Cyclotides written by and published by Academic Press. This book was released on 2015-11-24 with total page 404 pages. Available in PDF, EPUB and Kindle. Book excerpt: Advances in Botanical Research publishes in-depth and up-to-date reviews on a wide range of topics in plant sciences. Currently in its 76th volume, the series features several reviews by recognized experts on all aspects of plant genetics, biochemistry, cell biology, molecular biology, physiology and ecology. Publishes in-depth and up-to-date reviews on a wide range of topics in plant sciences Contains commentary by recognized experts on all aspects of plant genetics, biochemistry, cell biology, molecular biology, physiology, and ecology This volume features reviews of the fast moving field of plant cyclotides
Download or read book Fmoc Solid Phase Peptide Synthesis written by W. Chan and published by OUP Oxford. This book was released on 1999-12-16 with total page 371 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the publication of Atherton and Sheppard's volume, the technique of Fmoc solid-phase peptide synthesis has matured considerably and is now the standard approach for the routine production of peptides. The focus of this new volume is much broader, and covers the essential procedures.
Download or read book Chemical and Biological Synthesis written by Nick J Westwood and published by Royal Society of Chemistry. This book was released on 2018-08-16 with total page 317 pages. Available in PDF, EPUB and Kindle. Book excerpt: Synthetic chemistry plays a central role in many areas of chemical biology; utilising recent case studies, the goal of Chemical and Biological Synthesis is to highlight the full impact that the preparation of novel reagents can have in chemical biology. Covering the synthetic approaches that can be applied across the whole field of chemical biology, this book provides synthetic chemists with the broader context to which their work contributes and the biological questions that can be addressed through it. An ideal guide for postgraduate students and researchers in synthetic organic chemistry and chemical biology, Chemical and Biological Synthesis introduces synthetic techniques and methods to those who wish to incorporate synthesis for the first time in their biology-focused research programmes.
Download or read book Split Inteins written by Henning D. Mootz and published by Humana. This book was released on 2016-10-07 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume focuses on applications of split inteins, and the progress that has been made in the past 5 years on discovery and engineering of fast and more efficient split inteins. The first few chapters in Split Inteins: Methods and Protocols explore new techniques on how to use split inteins for affinity purification of overproduced proteins, and split-intein based technologies to prepare cyclic peptides and proteins. The next few chapters discuss semisynthetic protein trans-splicing using one synthetic intein piece, synthetic intein-extein pieces used to deliver other cargos for chemical modification both of purified proteins and of proteins in living cells, as well as isotopic labeling of proteins for NMR studies, and a discussion on how protein block copolymers can be generated by protein trans-splicing to form protein hydrogels. The last few chapters deal with intein applications in transgenic plants and conditional inteins that can be regulated in artificial ways by small molecules or light, a cassette-based approach to quickly test many intein insertion positions, and a computational approach to predict new intein split sites (the approach also works for other proteins). Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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. Cutting-edge and thorough, Split Inteins: Methods and Protocols is a valuable resource that will provide guidance toward possibilities of split intein applications, explore proven and detailed protocols adaptable to various research projects, and inspire new method developments.
Download or read book Expressed Protein Ligation written by Miquel Vila-Perelló and published by Humana. This book was released on 2020-03-07 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book provides a comprehensive overview of Expressed Protein Ligation (EPL), detailing methods and protocols to generate site-specifically modified proteins. Chapters include an overview of the protein semi-synthesis field, as well as related areas that have contributed to the development of EPL such as protein splicing and peptide synthesis. Following the introductory chapters, the rest of the book guides readers through protocols to perform EPL reactions, methods to synthesize peptide thioesters and to perform peptide and protein ligations, label proteins inside living cells, protocols for the semi-synthesis of phorphorylated, glycosylated and ubiquitylated proteins, synthesis and assembly of assymetrically modified nucleosomes, use of ligation auxiliaries and synthesis of cyclic proteins, as well as novel desulfurization strategies and use of selective Cys side chain protection to obtain precisely modified proteins.Written in the highly successful Methods in Molecular Biology 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. Authoritative and cutting-edge, Expressed Protein Ligation: Methods and Protocols will ensure successful implementation of protein semi-synthesis methods to further study the structure and function of proteins.
Download or read book Development of Chemistry Based Screening Platform for Access to Mirror Image Library of Natural Products written by Taro Noguchi and published by Springer. This book was released on 2017-11-03 with total page 89 pages. Available in PDF, EPUB and Kindle. Book excerpt: This thesis mainly describes the development of a screening process for a mirror-image library of chiral natural products. It demonstrates how, by using mirror-image proteins for the screening of available natural products, unavailable mirror-image isomers of natural products can be screened in a mirror process. Moreover, as mirror-image isomers including target proteins and natural products are mainly prepared by means of chemical synthesis, the screening strategy presented here suggests the importance of organic chemistry. Natural products are commonly used as valuable resources for drug discovery. However, as they are mostly produced as single enantiomeric forms, researchers have tested o nly natural products bearing one stereochemistry available in nature. As natural products and their enantiomers have identical physicochemical properties and different biological activities, mirror-image isomers of natural products are promising candidates for novel medicinal resources. In an effort to identify anticancer agents from the mirror-image library, chemical protein syntheses of some target oncoproteins, MDM2, MDMX and Grb2, and their applications to the chemical array screening process were achieved. In the course of this process the NP843 enantiomer, which is the enantiomer of an α-tocopherol derivative, was successfully identified as a novel MDM2-p53 interaction inhibitor. These results clearly show that a mirror-image library of chiral natural products represents an invaluable medicinal resource. Accordingly, the chemistry-based screening strategy described in this thesis will be of great interest to a broad range of chemists involved in natural product, medicinal, and synthetic chemistry.
Download or read book Bioconjugate Techniques written by Greg T. Hermanson and published by Academic Press. This book was released on 2010-07-26 with total page 1233 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioconjugate Techniques, 2nd Edition, is the essential guide to the modification and cross linking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or cross linking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. A one-stop source for proven methods and protocols for synthesizing bioconjugates in the lab Step-by-step presentation makes the book an ideal source for researchers who are less familiar with the synthesis of bioconjugates More than 600 figures that visually describe the complex reactions associated with the synthesis of bioconjugates Includes entirely new chapters on the latest areas in the field of bioconjugation as follows: Microparticles and nanoparticlesSilane coupling agentsDendrimers and dendronsChemoselective ligationQuantum dotsLanthanide chelatesCyanine dyesDiscrete PEG compoundsBuckyballs,fullerenes, and carbon nanotubesMass tags and isotope tagsBioconjugation in the study of protein interactions
Download or read book Wiley Encyclopedia of Chemical Biology Volume 1 written by Tadhg P. Begley and published by Wiley-Blackwell. This book was released on 2009 with total page 756 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first major reference at the interface of chemistry, biology, and medicine Chemical biology is a rapidly developing field that uses the principles, tools, and language of chemistry to answer important questions in the life sciences. It has enabled researchers to gather critical information about the molecular biology of the cell and is the fundamental science of drug discovery, playing a key role in the development of novel agents for the prevention, diagnosis, and treatment of disease. Now students and researchers across the range of disciplines that use chemical biology techniques have a single resource that encapsulates what is known in the field. It is an excellent place to begin any chemical biology investigation. Major topics addressed in the encyclopedia include: Applications of chemical biology Biomolecules within the cell Chemical views of biology Chemistry of biological processes and systems Synthetic molecules as tools for chemical biology Technologies and techniques in chemical biology Some 300 articles range from pure basic research to areas that have immediate applications in fields such as drug discovery, sensor technology, and catalysis. Novices in the field can turn to articles that introduce them to the basics, whereas experienced researchers have access to articles exploring the cutting edge of the science. Each article ends with a list of references to facilitate further investigation. With contributions from leading researchers and pioneers in the field, the Wiley Encyclopedia of Chemical Biology builds on Wiley's unparalleled reputation for helping students and researchers understand the crucial role of chemistry and chemical techniques in the life sciences.
Download or read book Peptide Applications in Biomedicine Biotechnology and Bioengineering written by Sotirios Koutsopoulos and published by Woodhead Publishing. This book was released on 2017-11-16 with total page 655 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peptide Applications in Biomedicine, Biotechnology and Bioengineering summarizes the current knowledge on peptide applications in biomedicine, biotechnology and bioengineering. After a general introduction to peptides, the book addresses the many applications of peptides in biomedicine and medical technology. Next, the text focuses on peptide applications in biotechnology and bioengineering and reviews of peptide applications in nanotechnology. This book is a valuable resource for biomaterial scientists, polymer scientists, bioengineers, mechanical engineers, synthetic chemists, medical doctors and biologists. Presents a self-contained work for the field of biomedical peptides Summarizes the current knowledge on peptides in biomedicine, biotechnology and bioengineering Covers current and potential applications of biomedical peptides
Download or read book Protein Self Assembly written by Jennifer J. McManus and published by Humana. This book was released on 2020-08-08 with total page 266 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores experimental and computational approaches to measuring the most widely studied protein assemblies, including condensed liquid phases, aggregates, and crystals. The chapters in this book are organized into three parts: Part One looks at the techniques used to measure protein-protein interactions and equilibrium protein phases in dilute and concentrated protein solutions; Part Two describes methods to measure kinetics of aggregation and to characterize the assembled state; and Part Three details several different computational approaches that are currently used to help researchers understand protein self-assembly. Written in the highly successful Methods in Molecular Biology 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. Thorough and cutting-edge, Protein Self-Assembly: Methods and Protocols is a valuable resource for researchers who are interested in learning more about this developing field.
