Engineering the Genetic Code and Expanding the Genetic Code for in Vivo Incorporation of Noncanonical Amino Acids Into Biologically Active Ribosomal Peptides

Engineering the Genetic Code and Expanding the Genetic Code for in Vivo Incorporation of Noncanonical Amino Acids Into Biologically Active Ribosomal Peptides PDF Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 95

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Non-Natural Amino Acids

Non-Natural Amino Acids PDF Author:
Publisher: Academic Press
ISBN: 0080921639
Category : Science
Languages : en
Pages : 334

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Book Description
By combining the tools of organic chemistry with those of physical biochemistry and cell biology, Non-Natural Amino Acids aims to provide fundamental insights into how proteins work within the context of complex biological systems of biomedical interest. The critically acclaimed laboratory standard for 40 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. With more than 400 volumes published, each Methods in Enzymology volume presents material that is relevant in today's labs -- truly an essential publication for researchers in all fields of life sciences. - Demonstrates how the tools and principles of chemistry combined with the molecules and processes of living cells can be combined to create molecules with new properties and functions found neither in nature nor in the test tube - Presents new insights into the molecular mechanisms of complex biological and chemical systems that can be gained by studying the structure and function of non-natural molecules - Provides a "one-stop shop" for tried and tested essential techniques, eliminating the need to wade through untested or unreliable methods

Cyclic Peptides

Cyclic Peptides PDF Author: Jesko Koehnke
Publisher: Royal Society of Chemistry
ISBN: 1788013778
Category : Science
Languages : en
Pages : 392

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Book Description
Cyclic peptides are increasingly employed as chemical tools in biology and drug discovery. They have gained a lot of interest as alternative sources of new drugs to traditional small molecules. This book introduces cyclic peptides and provides a thorough overview of biosynthetic and fully synthetic approaches to their preparation. Following an introduction to cyclic peptides, biosynthetic and traditional chemical routes to cyclic peptides are reviewed. Due to their size, their synthesis is not trivial. Recent advances in the incorporation of novel structural units are presented in addition to how synthesis and biological methods can be combined. The chemical analysis of this molecular class is also discussed. Furthermore, chapters detail the progression of cyclic peptides as tools in biology and as potential drugs, providing a future vision of their importance. In total, this book provides the reader with a comprehensive view of the state-of-the-art of cyclic peptides, from construction to possible clinical utility. This book will be an essential resource for students, researchers and scientists within industry in medicinal, bioorganic, natural product and analytical chemistry fields.

Unnatural Amino Acid Incorporation for Genetic Code Expansion in Mammalian Cells

Unnatural Amino Acid Incorporation for Genetic Code Expansion in Mammalian Cells PDF Author: Jeffrey Kunio Takimoto
Publisher:
ISBN: 9781124803968
Category :
Languages : en
Pages : 150

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Book Description
The genetic code of most organisms was evolved to encode 20 amino acids. Although the ability to encode 20 amino acids provides the basis to translate proteins necessary for life, researchers are also limited to these 20 amino acids for conventional site-directed mutagenesis. The ability to encode unnatural amino acids provides researchers the ability to circumvent the limitation imposed by the genetic code. Genetically encoding unnatural amino acids provides researchers the means to not only mimic naturally occurring posttranslational modifications but also the ability to encode amino acids with new physical or chemical properties to study biological processes. The incorporation of unnatural amino acids into proteins had been developed in Escherichia coli and also in yeast. We have developed a methodology to genetically incorporate unnatural amino acids in mammalian cells in response to an amber codon (UAG). The incorporation of unnatural amino acids is high in E. coli and yeast, but the incorporation in mammalian cells is relatively low. In addition to developing the system to incorporate unnatural amino acids in mammalian cells, we have also improved suppression efficiencies by modifying the synthetase and unnatural amino acid. To incorporate unnatural amino acids in response to an amber codon, the tRNA anticodon is mutated from a GUA to a CUA. We were able to show that engineering the anticodon-binding domain of the synthetase could enhance the recognition of the tRNA and thus increased suppression efficiencies. Furthermore, by masking the carboxyl group of the amino acid by an ester group, we were able to increase the bioavailability of an unnatural amino acid to further increase suppression efficiencies. Most evolved synthetases aminoacylate unnatural amino acids that are structurally similar to the native substrate of the wild-type synthetase. We were able to adapt Methanosarcina mazei pyrrolysine synthetase (PylRS) to charge a considerable disparate amino acid from its native substrate, O-methyl-L-tyrosine. In addition, the X-ray crystal structure was solved for the evolved PylRS complexed with O-methyl-L-tyrosine at 1.75Å. This multifaceted approach provides the basis to engineer the PylRS to incorporate a significantly diverse selection of unnatural amino acids than previously anticipated.

Engineering the Genetic Code

Engineering the Genetic Code PDF Author: Nediljko Budisa
Publisher: John Wiley & Sons
ISBN: 3527607099
Category : Science
Languages : en
Pages : 312

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Book Description
The ability to introduce non-canonical amino acids in vivo has greatly expanded the repertoire of accessible proteins for basic research and biotechnological application. Here, the different methods and strategies to incorporate new or modified amino acids are explained in detail, including a lot of practical advice for first-time users of this powerful technique. Novel applications in protein biochemistry, genomics, biotechnology and biomedicine made possible by the expansion of the genetic code are discussed and numerous examples are given. Essential reading for all molecular life scientists who want to stay ahead in their research.

Ancestral Sequence Reconstruction

Ancestral Sequence Reconstruction PDF Author: David A Liberles
Publisher: Oxford University Press
ISBN: 0199299188
Category : Science
Languages : en
Pages : 267

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Book Description
Ancestral sequence reconstruction is a technique of growing importance in molecular evolutionary biology and comparative genomics. As a powerful tool for testing evolutionary and ecological hypotheses, as well as uncovering the link between sequence and molecular phenotype, there are potential applications in a range of fields.Ancestral Sequence Reconstruction starts with a historical overview of the field, before discussing the potential applications in drug discovery and the pharmaceutical industry. This is followed by a section on computational methodology, which provides a detailed discussion of the available methods for reconstructing ancestral sequences (including their advantages, disadvantages, and potential pitfalls). Purely computational applications of the technique are then covered, including wholeproteome reconstruction. Further chapters provide a detailed discussion on taking computationally reconstructed sequences and synthesizing them in the laboratory. The book concludes with a description of the scientific questions where experimental ancestral sequence reconstruction has been utilized toprovide insights and inform future research.This research level text provides a first synthesis of the theories, methodologies and applications associated with ancestral sequence recognition, while simultaneously addressing many of the hot topics in the field. It will be of interest and use to both graduate students and researchers in the fields of molecular biology, molecular evolution, and evolutionary bioinformatics.

Hyperthermophilic Enzymes

Hyperthermophilic Enzymes PDF Author:
Publisher: Academic Press
ISBN: 9780121822323
Category : Science
Languages : en
Pages : 494

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Book Description
The critically acclaimed laboratory standard for more than forty years, Methods in Enzymologyis one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences. This volume and its companions (Volumes 330 and 334) cover all current knowledge concerning hyperthermophilic enzymes. Major topics in this volume include the enzymes of central metabolism and respiratory enzymes.

Synthetic Biology

Synthetic Biology PDF Author: Anton Glieder
Publisher: Springer
ISBN: 3319227084
Category : Science
Languages : en
Pages : 371

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Book Description
This book provides a comprehensive, up-to-date overview of the opportunities and challenges of the complex field of synthetic biology, which combines various scientific disciplines. The emerging field of synthetic biology employs biotechnological approaches to recreate and enhance basic biological structures, intracellular processes and whole organisms. The book addresses a broad range of topics, including redesigning complex metabolic pathways, DNA/RNA and protein engineering, as well as novel synthetic biomaterials. It discusses both “bottom up” and “top down” approaches and presents the latest genome engineering tools with predictions about how these could change our way of thinking and working. Since the use of synthetic biology raises a number of ethical questions, a chapter is devoted to public awareness and risk management.The book is of interest to scientists from both academia and industry, as well as PhD students and postdocs working in the field

Recoding: Expansion of Decoding Rules Enriches Gene Expression

Recoding: Expansion of Decoding Rules Enriches Gene Expression PDF Author: John F. Atkins
Publisher: Springer Science & Business Media
ISBN: 0387893822
Category : Science
Languages : en
Pages : 473

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Book Description
The literature on recoding is scattered, so this superb book ?lls a need by prov- ing up-to-date, comprehensive, authoritative reviews of the many kinds of recoding phenomena. Between 1961 and 1966 my colleagues and I deciphered the genetic code in Escherichia coli and showed that the genetic code is the same in E. coli, Xenopus laevis, and guinea pig tissues. These results showed that the code has been c- served during evolution and strongly suggested that the code appeared very early during biological evolution, that all forms of life on earth descended from a c- mon ancestor, and thus that all forms of life on this planet are related to one another. The problem of biological time was solved by encoding information in DNA and retrieving the information for each new generation, for it is easier to make a new organism than it is to repair an aging, malfunctioning one. Subsequently, small modi?cations of the standard genetic code were found in certain organisms and in mitochondria. Mitochondrial DNA only encodes about 10–13 proteins, so some modi?cations of the genetic code are tolerated that pr- ably would be lethal if applied to the thousands of kinds of proteins encoded by genomic DNA.

Peptide Macrocycles

Peptide Macrocycles PDF Author: Matthew B. Coppock
Publisher: Humana
ISBN: 9781071616888
Category : Technology & Engineering
Languages : en
Pages : 469

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Book Description
This volume explores the latest techniques and strategies used to study the field of peptide macrocycles. The chapters in this book ae organized into four parts: macrocycles synthesis, combinational library synthesis and screening, macrocycle characterization, and unique applications. Part One looks at a variety of peptide cyclization methodologies, and Part Two describes methods for the creation of peptide macrocycles libraries and their subsequent screening against biological targets of interest. Part Three discusses the study and characterization of peptide macrocycle-target interactions, and Part Four introduces unique applications for peptide macrocycles, from higher-order structure formation to post-synthetic functional modifications. 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. Cutting-edge and comprehensive, Peptide Macrocycles: Methods and Protocols is a valuable resource for both novice and expert researchers looking to learn more about this developing field.