Analytical ultracentrifugation (AUC) can supply rich information on the mass, shape, size distribution, solvation, and composition of macromolecules and nanoscopic particles. It also provides a detailed view of their reversible single- or multi-component interactions over a wide range of affinities. Yet this powerful technique has been hard to mast
Analytical ultracentrifugation has become an increasingly important technique for monitoring the size and shape of biological macromolecules. Analytical Ultracentrifugation: Techniques and Methods contains contributions from experts in the field, bringing together the multitude of developments that have taken place in instrumentation and analysis over the past decade into a single volume. This book covers the latest methods in analysis along with an extensive introduction for the novice user. Analysis methods in both sedimentation velocity and sedimentation equilibrium are discussed at length. Protein, protein/DNA, membrane proteins and polymer systems are also explored, along with software developments and non-ideality.
This volume successfully and clearly examines how biophysical approaches can be used to study complex systems of reversibly interacting proteins. It deals with the methodology behind the research and shows how to synergistically incorporate several methodologies for use. Each chapter treats and introduces the reader to different biological systems, includes a brief summary of the physical principles, and mentions practical requirements.
There are numerous examples in the history of science when the parallel develop ments of two or more disciplines, methodologies, technologies or theoretical in sights have converged to produce significant scientific advances. The decades following the 1950s have produced several such significant advances, as a result of a convergence of developments in molecular biology and in solid state-based electronics instrumentation. Since one of these areas of significant advancement, analytical ultracentrifu gation, has been undergoing a renaissance, we thought it would be a useful activity to call upon a group of researchers who have been developing either the experi mental or theoretical aspects of the methodology and gather in one place a group of articles summarizing the current status of the field. The success of recombinant DNA methodologies at producing biologically active macromolecules of commer cial interest has evoked interests in mechanisms of function. Pursuit of the related questions has emphasized the importance of studies of macromolecular binding and interaction. Several contributions to this volume remind us that analytical ultra centrifugation is rigorously based on solid thermodynamic theory and, as such, is fully capable of providing comprehensive quantitative descriptions of molecular interactions in solution. Furthermore, a number of the chapters provide examples, along with innovative methods for carrying out these characterizations. The past decade has seen several developments that reflect the rebirth of interest in analytical ultracentrifugation.
This is the first book of its kind to appear for nearly two decades and gives as comprehensive a coverage as is possible of the present state-of-the-art.
Ultracentrifugation in Biochemistry discusses the fundamental aspects of ultracentrifugation. The book begins with a sketch of the field, highlighting some of the principal developments. Following this is a chapter that discusses ultracentrifugation in general terms and describes the division of the field into three major areas. The subsequent chapter deals with developments of the experimental aspects of the field such as improvements in the instrument itself, cells, rotors, measurement, and control of temperature, and the various optical systems. The remainder of the book discusses the fundamental principles of sedimentation velocity, transient states, and sedimentation equilibrium. A section is also included which deals with interpretation of sedimentation data in terms of hydrodynamic models, charge effects, and interactions in multicomponent systems. This book is likely to become an indispensable companion to the laboratory worker who is planning and conducting an ultracentrifuge run for almost any purpose. It should also be of fundamental value to the thoughtful student or investigator who wants to know the present state of knowledge in the field, both experimental and theoretical.
This book introduces analytical ultracentrifugation (AUC) as a whole, covering essential theoretical and practical aspects as well as its applications in both biological and non-biological systems. Comprehensive characterizations of macromolecules in a solution are now routinely required not only for understanding the solution system but also for producing a solution with better properties. Analytical ultracentrifugation is one of most powerful and reliable techniques for studying the biophysical behavior of solutes in solution. In the last few years, there have been steady advances made in hardware, software, and applications for AUC. This book provides chapters that cover everything essential for beginners to the most advanced users and also offer updated knowledge of the field on advances in hardware, software, and applications. Recent development of hardware described in this book covers new detection systems that give added dimensions to AUC. Examples of data analysis with essential theoretical explanations for advanced and recently updated software are also introduced. Besides AUC of biological systems including membrane proteins and biopharmaceuticals, AUC applications for non-biological questions are included. AUC studies under non-ideal conditions such as highly concentrated solutions and solutions with high salt concentration are also included. The contributors to this book are leading researchers in the fields of solution biophysics and physical chemistry who extensively employ AUC analysis for their research. From this published work, one can gain new and comprehensive knowledge of recent AUC analysis.
Centrifugal Separations in Molecular and Cell Biology focuses on the application of modern centrifugation technology in molecular and cell biology, including the separation and fractionation of biological particles by centrifugation on the preparative and analytical scales. The selection first covers the principles and practices of centrifugation and the bases of centrifugal separations. Discussions focus on the basic concepts of sedimentation theory, centrifugation methods, designing centrifugation experiments, care of centrifuges and rotors, and statistical estimation of molecular parameters. The book also ponders on the practical aspects of rate-zonal centrifugation, including gradient materials, density and viscosity of glycerol solutions, and resolution and gradient shape. The publication examines fractionations in zonal rotors and the quantitative aspects of rate-zonal centrifugation. The text then reviews isopycnic centrifugation in ionic media and analytical centrifugation. Topics include separation by isopycnic banding, large-scale preparative procedures, and density-gradient solutes. The selection is a valuable reference for readers interested in centrifugation technology.
Mathematical Theory of Sedimentation Analysis deals with ultracentrifugal analysis. The book reviews flow equations for the ultracentrifuge, for two component systems, for multicomponent systems, and in chemically reacting systems. It explains the Svedberg equation and its extensions, and also the tests of the Onsager reciprocal relation. By employing a system consisting of two strong electrolytes and a solvent, the book illustrates that the sedimentation processes can be treated in terms of thermodynamics of irreversible processes. It also explains sedimentation-diffusion equilibrium and an approach to sedimentation equilibrium. It reviews the prediction of the time required to reach equilibrium, the estimates being made by Weaver (1926), and by Mason and Weaver (1924). The book employs sedimentation in a sector-shaped cell in a centrifugal field, of which the solutions of Mason and Weaver closely approximate the actual concentration distribution in the ultra-centrifuge cell. Other accurate solutions are by Fujita, Nazarian (1958), Yphantis, and Waugh. The book will prove valuable for mathematicians, physical chemists, biophysical chemists students, or professor of advanced mathematics.
The book is structured in nine sections, each containing several chapters. The volume starts with an overview of analytical techniques and progresses through purification of proteins; protein modification and inactivation; protein size, shape, and structure; enzyme kinetics; protein-ligand interactions; industrial enzymology; and laboratory quality control. The book is targeted at all scientists interested in protein research.
