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The metabolism of all living organisms consists of elaborate and complex interactions among numbers of biomolecules, including protein-protein interactions. BIACORE is a new device that uses surface plasmon resonance for kinetic analysis of molecular interactions and automates whole analytical processes. The number of users of BIACORE is increasing worldwide, but until now nothing had been published in English to explain the principles of this new methodology and the kinds of applications it makes available. This updated and revised version of the Japanese original explains the underlying principles of BIACORE, providing concrete examples that utilize the methodology for analysis in areas ranging from basic to applied sciences. With its generous use of illustrations, this book is a valuable source of information for all users of BIACORE.
Biomolecular Interactions: Part A, Volume 169, the latest release in the Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of timely topics, including Emerging Mechanisms of Targeted Protein Degradation by Molecular Glues, Design and use of programmable DNA Hydrogels, Oligomerization of membrane receptors: Approaches to measure in live cells, Interactions of alpha-synuclein with biomolecules, Gel-electrophoresis based method for biomolecular interaction, Recombinant centrosome expression in bacterial system, Reconstituting CCL5-CCR5 complex for structural and mechanistic analysis, Protein engineering and design in ion channel receptors, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Cell Biology series - Updated release includes the latest information on biomolecular interactions instead of protein-protein interactions
With its exploration of the scientific and technological characteristics of systems exploiting molecular recognition between synthetic materials, such as polymers and nanoparticles, and biological entities, this is a truly multidisciplinary book bridging chemistry, life sciences, pharmacology and medicine. The authors introduce innovative biomimetic chemical assemblies which constitute platforms for recruitment of cellular components or biological molecules, while also focusing on physical, chemical, and biological aspects of biomolecular recognition. The diverse applications covered include biosensors, cell adhesion, synthetic receptors, cell patterning, bioactive nanoparticles, and drug design.
Surface plasmon resonance (SPR) plays a dominant role in real-time interaction sensing of biomolecular binding events, this book provides a total system description including optics, fluidics and sensor surfaces for a wide researcher audience.
"The characterization of biomolecular interactions is an important step towards understanding the basic biochemical processes that govern cellular function, the aberrations that cause disease and potential therapeutic candidates. The goal of my thesis research has been to develop Arrayed Imaging Reflectometry (AIR) as a technique to characterize biomolecular interactions in terms of their binding affinity and binding kinetics. AIR is an interferometric biosensing technique that relies on the creation of a total destructive interference condition on a silicon chip coated with a thin layer of probe-functionalized silicon dioxide. Specific binding of target molecules results in an increase in the local thickness of the probe layer and a concomitant destruction of this total destructive interference condition, resulting in an increased reflectance that corresponds to the amount of bound target. Initial studies towards this goal focused on the optimization of surface functionalization chemistry for AIR experiments. Next, experiments were carried out to confirm the quantitative performance of AIR across a range of probe-analyte affinities. We combined biomolecular binding models with data obtained from spectroscopic ellipsometry and surface plasmon resonance experiments to model AIR response for two probe-analyte pairs and compared it to experimental observations. This study confirmed that AIR response is a predictable function of probe and analyte properties and thus could be used to determine binding parameters for biomolecular interactions. A modified substrate consisting of a silicon nitride/ silicon dioxide bilayer on a silicon chip was introduced to enable AIR detection under aqueous media. The thickness sensitivity of this substrate was experimentally determined to be 1 Å. We used the new substrate to measure the binding kinetics of Muscleblind-like 1 (MBNL1), a splicing regulator protein that is implicated in the pathology of the Myotonic Dystrophies and Huntington Disease, to several of its 'normal' and 'pathogenic' RNA targets microarrayed on an AIR chip. The binding kinetics of 'normal' and 'pathogenic' RNA sequences were observed to be distinctly different even though their steady state binding constants were similar. These results suggest that AIR can be a powerful technique for the label-free real-time analysis of biomolecular interactions in a high-throughput format"--Pages viii-ix.
Hands-on researchers describe in step-by-step detail 73 proven laboratory methods and bioinformatics tools essential for analysis of the proteome. These cutting-edge techniques address such important tasks as sample preparation, 2D-PAGE, gel staining, mass spectrometry, and post-translational modification. There are also readily reproducible methods for protein expression profiling, identifying protein-protein interactions, and protein chip technology, as well as a range of newly developed methodologies for determining the structure and function of a protein. The bioinformatics tools include those for analyzing 2D-GEL patterns, protein modeling, and protein identification. All laboratory-based protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Biomolecular Interactions: Part A, Volume 166, the latest release in the Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of timely topics in cell biology. Each chapter is written by an international board of authors. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Cell Biology series - Updated release includes the latest information on biomolecular interactions instead of protein-protein interactions
The development of devices that incorporate biological assemblies is impacting analytical and biomedical research. Today, scientists can monitor vital biological interactions-such as the binding of DNA to proteins-in real time, deriving unique information necessary to understanding biochemical pathways and thus aiding the design of drugs to regulat