Enzyme Mediated Ligation Methods
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Enzyme Mediated Ligation Methods
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Author : Timo Nuijens
language : en
Publisher: Springer Nature
Release Date : 2019-06-03
Enzyme Mediated Ligation Methods written by Timo Nuijens and has been published by Springer Nature this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019-06-03 with Science categories.
This volume discusses different enzyme-catalyzed ligation methodologies for a variety of different chemical transformations. This book wants readers to view enzymes as a powerful tool in both academic and industrial research. Chapters in this book cover topics such as sortase A-mediated generation of site-specifically conjugated antibody-drug conjugates; omniligase-catalyzed inter- and intramolecular ligation; ligation catalyzed by microbial transglutaminase; peptide cyclization mediated by cyanobactin macrocyclases, butelase 1 and sortase A; using BioID as a tool for protein proximity labeling in living cells; and inducible, selective labeling of proteins via enzymatic oxidation of tyrosine. 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 thorough, Enzyme-Mediated Ligation Methods is a valuable resource for students and scientists from different disciplines who are interested in using enzymatic strategies to answer their research questions.
Enzyme Mediated Labeling Of Proteins And Protein Protein Interactions In Vitro And In Living Cells
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Author : Sarah Ann Slavoff
language : en
Publisher:
Release Date : 2010
Enzyme Mediated Labeling Of Proteins And Protein Protein Interactions In Vitro And In Living Cells written by Sarah Ann Slavoff and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010 with categories.
The E. coli biotin ligase enzyme, BirA, has been previously used by the Ting research group for site-specific labeling of peptide-tagged cell surface proteins. We sought to expand the utility of biotin ligase-mediated labeling to functional group handles, including azides and alkynes, for bio-orthogonal chemistry. Since the BirA and its point mutants were unable to ligate these probes to an acceptor peptide, we screened biotin ligases from multiple species to identify more permissive enzymes. We determined that the Pyrococcus horikoshii biotin ligase utilizes an azide-bearing biotin analog and that the Saccharomyces cerevisiae biotin ligase can utilize an alkyne-functionalized biotin analog. We subsequently demonstrated that the azidefunctionalized biotin analog can be derivatized with a phosphine probe via the Staudinger ligation. We next turned to the goal of delivering quantum dots to the cytosol of living cells, which in the future may permit intracellular single-molecule imaging. We investigated viral methods of delivery, but found that our protocol caused quantum dots to be trapped in endocytic vesicles. We then validated previous reports that the pore-forming toxin streptolysin 0 be used to deliver quantum dots to the cytosol of living cells. Lipoic acid ligase, or LpIA, has been previously applied to site-specific protein labeling of peptide-tagged proteins using small molecule probes including lipoic acid and coumarin fluorophores. We utilized LpIA and its substrate, the LAP peptide, to create sensors for proteinprotein interactions. If LpIA is fused to one protein and LAP is fused to another, only when the two proteins interact do LpIA and LAP come into proximity, allowing probe ligation onto the peptide to occur as a readout of the interaction. We demonstrate that proximity-dependent coumarin ligation detects protein-protein interactions in living mammalian cells with extremely low background, a signal-to-background ratio of at least 5:1, and sufficiently fast kinetics to label interactions with a half-life of at least 1 minute. The reporter quantitatively responds to subpopulations of interacting proteins, allowing dissociation constants to be measured. Coumarin fluorescence accurately reports the subcellular localization of the interaction under study. Finally, we applied proximity-dependent coumarin ligation to imaging of the interaction of PSD-95 and neuroligin-1, two proteins involved in synaptic maturation, in neurons.
Chemoselective And Bioorthogonal Ligation Reactions
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Author : W. Russ Algar
language : en
Publisher: John Wiley & Sons
Release Date : 2017-06-19
Chemoselective And Bioorthogonal Ligation Reactions written by W. Russ Algar and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017-06-19 with Technology & Engineering categories.
This timely, one-stop reference is the first on an emerging and interdisciplinary topic. Covering both established and recently developed ligation chemistries, the book is divided into two didactic parts: a section that focuses on the details of bioorthogonal and chemoselective ligation reactions at the level of fundamental organic chemistry, and a section that focuses on applications, particularly in the areas of chemical biology, biomaterials, and bioanalysis, highlighting the capabilities and benefits of the ligation reactions. With chapters authored by outstanding scientists who range from trailblazers in the field to young and emerging leaders, this book on a highly interdisciplinary topic will be of great interest for biochemists, biologists, materials scientists, pharmaceutical chemists, organic chemists, and many others.
Chemoselective And Bioorthogonal Ligation Reactions
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Author : W. Russ Algar
language : en
Publisher: John Wiley & Sons
Release Date : 2017-03-17
Chemoselective And Bioorthogonal Ligation Reactions written by W. Russ Algar and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017-03-17 with Technology & Engineering categories.
This timely, one-stop reference is the first on an emerging and interdisciplinary topic. Covering both established and recently developed ligation chemistries, the book is divided into two didactic parts: a section that focuses on the details of bioorthogonal and chemoselective ligation reactions at the level of fundamental organic chemistry, and a section that focuses on applications, particularly in the areas of chemical biology, biomaterials, and bioanalysis, highlighting the capabilities and benefits of the ligation reactions. With chapters authored by outstanding scientists who range from trailblazers in the field to young and emerging leaders, this book on a highly interdisciplinary topic will be of great interest for biochemists, biologists, materials scientists, pharmaceutical chemists, organic chemists, and many others.
Organic Reactions Volume 97
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Author :
language : en
Publisher: John Wiley & Sons
Release Date : 2019-01-14
Organic Reactions Volume 97 written by and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019-01-14 with Science categories.
The 97th volume in this series for organic chemists in industry presents critical discussions of widely used organic reactions or particular phases of a reaction. The material is treated from a preparative viewpoint, with emphasis on limitations, interfering influences, effects of structure and the selection of experimental techniques. The work includes tables that contain all possible examples of the reaction under consideration. Detailed procedures illustrate the significant modifications of each method.
Proximity Labeling
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Author : Murat Sunbul
language : en
Publisher: Springer Nature
Release Date : 2019-05-23
Proximity Labeling written by Murat Sunbul and has been published by Springer Nature this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019-05-23 with Science categories.
This book provides detailed protocols and untold tips and tricks regarding the most well-known examples of proximity labeling methods, in which the protein of interest is genetically fused to or labeled with an enzyme that can generate short-lived reactive species to non-specifically label molecules within a certain radius of up to twenty nanometers. Beginning with peroxidase-based proximity labeling methods, the volume continues with BioID, proximity labeling methods that describe the proximity ligation assay to detect RNA-DNA interactions, UV cross-linking to demonstrate RNA-protein interactions, and how chemical and enzymatic reactivities can be improved upon DNA-DNA and protein-protein interactions, as well as “proximity-induced self-labeling,” where the radius of labeling is zero. Written for the highly successful Methods in Molecular Biology series, 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 practical, Proximity Labeling: Methods and Protocols serves as an ideal guide for researchers exploring the crucial roles that proximity-driven reactions play in biological systems.
Enzymes Of Epigenetics
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Author :
language : en
Publisher: Academic Press
Release Date : 2016-06-29
Enzymes Of Epigenetics written by and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2016-06-29 with Science categories.
Enzymes of Epigenetics, one of two new volumes in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods that are employed to study epigenetic regulation and includes structural, biochemical, molecular, biological, cellular, computational, and systems approaches. Topics include chromatin structure and histones, posttranslational histone modification enzymes and complexes, histone modification binders, DNA modifications and nucleic acid regulators, epigenetic technologies and small molecule epigenetic regulators, and biological connections Continues the legacy of this premier serial with quality chapters authored by leaders in the field Contains two new volumes covering research methods in enzymes of epigenetics Covers topics such as chromatin structure and histones, posttranslational histone modification enzymes and complexes, histone modification binders, DNA modifications and nucleic acid regulators, epigenetic technologies and small molecule epigenetic regulators and biological connections
Non Ribosomal Peptide Biosynthesis And Engineering
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Author : Michael Burkart
language : en
Publisher: Springer Nature
Release Date : 2023-05-15
Non Ribosomal Peptide Biosynthesis And Engineering written by Michael Burkart and has been published by Springer Nature this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023-05-15 with Science categories.
This volume provides new technologies on NRPSs and related carrier protein dependent synthases, including polyketide synthases (PKS) and fatty acid synthases (FAS). Chapters detail enzymology, structural biology, proteopromics, chemical biology, natural product chemistry, and bioinformatics. 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 methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Non-Ribosomal Peptide Biosynthesis and Engineering: Methods and Protocols aims to feature methods that will be beneficial to new researchers, and those wanting to adopt new methodologies into their research.
Plant Cyclotides
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Author :
language : en
Publisher: Academic Press
Release Date : 2015-11-24
Plant Cyclotides written by and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2015-11-24 with Science categories.
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
Engineering The Genetic Code
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Author : Nediljko Budisa
language : en
Publisher: John Wiley & Sons
Release Date : 2006-05-12
Engineering The Genetic Code written by Nediljko Budisa and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2006-05-12 with Science categories.
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.