Synthesis of Proteins by Native Chemical Ligation Using Fmoc-Based Chemistry PDF Download

Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 11

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Book Description
C-terminal peptide [alpha]-thioesters are valuable intermediates in the synthesis/semisynthesis of proteins by native chemical ligation. They are prepared either by solid-phase peptide synthesis (SPPS) or biosynthetically by protein splicing techniques. The present paper reviews the different methods available for the chemical synthesis of peptide [alpha]-thioesters using Fmoc-based SPPS.

Author: Xuechen Li
Publisher: Springer Nature
ISBN: 107162489X
Category : Science
Languages : en
Pages : 258

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Book Description
This volume provides updated protocols for chemical protein synthesis. Chapters guide readers through development methods, strategies, and applications of protein chemical synthesis. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Chemical Protein Synthesis aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

Author: Luca D. D'Andrea
Publisher: John Wiley & Sons
ISBN: 1119044138
Category : Science
Languages : en
Pages : 558

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Book Description
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

Author: Yinfeng Zhang
Publisher: Open Dissertation Press
ISBN: 9781361345481
Category :
Languages : en
Pages :

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Book Description
This dissertation, "Protein Chemical Synthesis by Serine and Threonine Ligation" by Yinfeng, Zhang, 张银凤, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Landmark advances in the field of synthetic protein chemistry have enabled the preparation of complex, homogeneous proteins, including those that carry specific posttranslational modifications (PTMs). In addition, chemical synthesis will allow one to incorporate unnatural elements to generate new biologics with altered properties and functions. Native chemical ligation (NCL) is a milestone in the chemical synthesis of proteins (Kent et al., Science, 1994, 266, 776-779), in which a C-terminal peptide thioester and an N-terminal cysteine (Cys)-containing peptide-both in side-chain unprotected forms-are selectively coupled to generate a natural peptidic linkage at the site of ligation. This method requires a cysteine at the optimal convergent ligation site. However, Cys is one of the least abundant amino acids in natural proteins. Therefore, the development of new ligation methods at other amino acids will be necessary and important in this regard. Along these lines, our laboratory has developed a novel thiol-independent approach-serine/threonine ligation (STL). It uses the N-terminal serine or threonine of a peptide segment to chemoselectively react with another peptide segment with a C-terminal salicylaldehyde ester to form an N, O-benzylidene acetal linked product, followed by acidolysis to afford the final product at the natural Ser/Thr site. To extend the application of STL in chemical protein synthesis, we have developed a robust method for the preparation of peptide salicylaldehyde esters via Fmoc-based solid phase peptide synthesis. Furthermore, we have successfully applied this ligation method in the convergent synthesis of peptide drugs of significant therapeutic importance, including Teriparatide (Forteo), Corticorelin (oCRH), Exenatide (Byetta) and Tesamorelin (hGHRH). Of significance, we have demonstrated the effectiveness of our STL in the assembly of a more complex target of biological interest: human erythrocyte acylphosphatase ( 11 kDa). In summary, we have developed a new serine/threonine ligation, which can be effectively used to synthesize peptides and proteins. As there are countless serine and threonine residues in natural proteins, particularly those carrying posttranslational modifications, this method is anticipated to offer new opportunities in synthetic protein chemistry and chemical biology. DOI: 10.5353/th_b5295527 Subjects: Proteins - Synthesis

Author: Lei Liu
Publisher: Springer
ISBN: 3319191896
Category : Science
Languages : en
Pages : 235

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Book Description
Each review within the volume critically surveys one aspect of that topic and places it within the context of the volume as a whole. The most significant developments of the last 5 to 10 years are presented using selected examples to illustrate the principles discussed. The coverage is not intended to be an exhaustive summary of the field or include large quantities of data, but should rather be conceptual, concentrating on the methodological thinking that will allow the non-specialist reader to understand the information presented. Contributions also offer an outlook on potential future developments in the field.

Author: Ashraf Brik
Publisher: John Wiley & Sons
ISBN: 3527346600
Category : Science
Languages : en
Pages : 626

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Book Description
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.

Author: Christina Nadler
Publisher: Cuvillier Verlag
ISBN: 3736945043
Category : Science
Languages : en
Pages : 198

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Book Description
There is a constant need for developing improved methods for introducing artificial functionalities into peptides and proteins, as the modification of peptides and proteins is one of the major routes to investigate biological function in vitro and in vivo, e.g. by introduction of spin labels or fluorophores. To improve the synthetic accessibility of chemically modified peptides and proteins a new cysteine-free Native Chemical Ligation strategy based on a photocleavable auxiliary was developed and successfully implemented. In addition, a novel protocol for labeling peptides and proteins by introducing artificial, histidine-mimicking amino acids was devised. These triazole-based building blocks were utilized for the introduction of additional metal binding sites into peptides as well as for the development of peptidic zinc sensors based on zinc finger peptide Zif268.

Author: Lei Liu
Publisher: Springer
ISBN: 3319191861
Category : Science
Languages : en
Pages : 275

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Book Description
Each review within the volume critically surveys one aspect of that topic and places it within the context of the volume as a whole. The most significant developments of the last 5 to 10 years are presented using selected examples to illustrate the principles discussed. The coverage is not intended to be an exhaustive summary of the field or include large quantities of data, but should rather be conceptual, concentrating on the methodological thinking that will allow the non-specialist reader to understand the information presented. Contributions also offer an outlook on potential future developments in the field.

Author: Miquel Vila-Perelló
Publisher: Humana
ISBN: 9781071604335
Category : Science
Languages : en
Pages : 0

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Book Description
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.

Author: Nigel Stokes
Publisher:
ISBN: 9781682865965
Category : Science
Languages : en
Pages : 223

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Book Description
Proteins are organic compounds which are formed of amino acids that are linked together by peptides. They help the body in getting nitrogen, vitamins and sulfur. Proteins are three dimensional in their structure. Their structure can be categorized into four distinctive aspects - primary structure, secondary structure, quaternary structure and tertiary structure. As this subject is emerging at a rapid pace, the contents of this book will help the readers understand the modern concepts and applications of the subject. This book is meant for students who are looking for an elaborate reference text on protein chemistry.