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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.
Recognition receptors play a key role in the successful implementation of chemical and biosensors. Molecular recognition refers to non-covalent speci?c binding between molecules, one of which is typically a macromolecule or a molecular assembly, and the other is the target molecule (ligand or analyte). Biomolecular recognition is typically driven by many weak interactions such as hydrogen bo- ing, metal coordination, hydrophobic forces, van der Waals forces, pi-pi interactions and electrostatic interaction (due to permanent charges, dipoles, and quadrupoles) the polarization of charge distributions by the interaction partner leading to ind- tion and dispersion forces, and Pauli-exclusion-principle-derived inter-atomic repulsion, and a strong, “attractive” force arising largely from the entropy of the solvent and termed the hydrophobic effect. In recent years, there has been much progress in understanding the forces that drive the formation of such complexes, and how these forces are relate to the physical properties of the interacting molecules and their environment allows rational design of molecules and materials that interact in speci?c and desired ways. This book presents a signi?cant and up-to-date review of the various recognition elements, their immobilization, characterization techniques by a panel of dist- guished scientists. This work is a comprehensive approach to the recognition receptors area presenting a thorough knowledge of the subject and an effective integration of these receptors on sensor surfaces in order to appropriately convey the state-of the-art fundamentals and applications of the most innovative approaches.
The use of light-emitting proteins for the detection of biomolecules provides fast and sensitive methods which overcome the disadvantages of radioactive labels and the high cost of fluorescent dyes. This reference work summarizes modern advanced techniques and their applications and includes practical examples of assays based on photoproteins. The book presents contemporary key topics like luminescent marine organisms, DNA probes, reporter gene assays and photoproteins, ratiometric sensing, use of photoproteins for in vivo functional imaging and luminescent proteins in binding assays, to name just a few, and is complemented by recent advances in instrumentation. Includes an introductory chapter by 2008 Chemistry Nobel laureate Osamu Shimomura.
In the last two decades, semiconductor quantum dots—small colloidal nanoparticles—have garnered a great deal of scientific interest because of their unique properties. Among nanomaterials, CdTe holds special technological importance as the only known II–VI material that can form conventional p–n junctions. This makes CdTe very important for the development of novel optoelectronic devices such as light-emitting diodes, solar cells, and lasers. Moreover, the demand for water-compatible light emitters and the most common biological buffers give CdTe quantum dots fields a veritable edge in biolabeling and bioimaging. Cadmium Telluride Quantum Dots: Advances and Applications focuses on CdTe quantum dots and addresses their synthesis, assembly, optical properties, and applications in biology and medicine. It makes for a very informative reading for anyone involved in nanotechnology and will also benefit those scientists who are looking for a comprehensive account on the current state of quantum dot–related research.
This volume provides a collection of contemporary perspectives on using activity-based protein profiling (ABPP) for biological discoveries in protein science, microbiology, and immunology. A common theme throughout is the special utility of ABPP to interrogate protein function and small-molecule interactions on a global scale in native biological systems. Each chapter showcases distinct advantages of ABPP applied to diverse protein classes and biological systems. As such, the book offers readers valuable insights into the basic principles of ABPP technology and how to apply this approach to biological questions ranging from the study of post-translational modifications to targeting bacterial effectors in host-pathogen interactions.
The series Topics in Current Chemistry presents critical reviews of the present and future trends in modern chemical research. The scope of coverage is all areas of chemical science including the interfaces with related disciplines such as biology, medicine and materials science. The goal of each thematic volume is to give the non-specialist reader, whether in academia or industry, a comprehensive insight into an area where new research is emerging which is of interest to a larger scientific audience. 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. Review articles for the individual volumes are invited by the volume editors. Readership: research chemists at universities or in industry, graduate students.
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.
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.