Microcalorimetry Of Biological Molecules

This volume provides methods on microcalorimetry approaches to investigate complex biological molecular systems. Chapters guide readers through Differential Scanning Calorimetry (DSC), Isothermal Titration Calorimetry (ITC), and advanced data processing. 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 practical, Microcalorimetry of Biological Molecules: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Examining the physical basis of the structure of macromolecules—proteins, nucleic acids, and their complexes—using calorimetric techniques Many scientists working in biology are unfamiliar with the basics of thermodynamics and its role in determining molecular structures. Yet measuring the heat of structural change a molecule undergoes under various conditions yields information on the energies involved and, thus, on the physical bases of the considered structures. Microcalorimetry of Macromolecules offers protein scientists unique access to this important information. Divided into thirteen chapters, the book introduces readers to the basics of thermodynamics as it applies to calorimetry, the evolution of the calorimetric technique, as well as how calorimetric techniques are used in the thermodynamic studies of macromolecules, detailing instruments for measuring the heat effects of various processes. Also provided is general information on the structure of biological macromolecules, proteins, and nucleic acids, focusing on the key thermodynamic problems relating to their structure. The book covers: The use of supersensitive calorimetric instruments, including micro and nano-calorimeters for measuring the heat of isothermal reactions (Isothermal Titration Nano-Calorimeter), the heat capacities over a broad temperature range (Scanning Nano-Calorimeter), and pressure effects (Pressure Perturbation Nano-Calorimeter) Two of the simplest but key structural elements: the α and polyproline helices and their complexes, the α-helical coiled-coil, and the pyroline coiled-coils Complicated macromolecular formations, including small globular proteins, multidomain proteins and their complexes, and nucleic acids Numerous examples of measuring the ground state of protein energetics, as well as changes seen when proteins interact The book also reveals how intertwined structure and thermodynamics are in terms of a macromolecule's organization, mechanism of formation, the stabilization of its three-dimensional structure, and ultimately, its function. The first book to describe microcalorimetric technique in detail, enough for graduate students and research scientists to successfully plumb the structural mysteries of proteins and the double helix, Microcalorimetry of Macromolecules is an essential introduction to using a microcalorimeter in biological studies.

This book is an up-to-date account of microcalorimetry as applied to biological topics and gives a comprehensive review of this growing area of research. For non- calorimetrists it is intended as an introduction and for those already involved in calorimetry it provides a survey of other important biological studies. The topics covered range from "pure" model biochemical systems to complex, intact, biological systems such as blood: also included are discussions of both practical and theoretical problems.

Volume 323 of Methods in Enzymology is dedicated to the energetics of biological macromolecules. Understanding the molecular mechanisms underlying a biological process requires detailed knowledge of the structural relationships within the system and an equally detailed understanding of the energetic driving forces that control the structural interactions. This volume presents modern thermodynamic techniques currently being utilized to study the energetic driving forces in biological systems. It will be a useful reference source and textbook for scientists and students whose goal is to understand the energetic relationships between macromoleculer structures and biological functions. This volume supplements Volumes 259 and Volume 295 of Methods in Enzymology. Key Features * Probing Stability of Helical Transmembrane Proteins * Energetics of Vinca Alkaloid Interactions with Tubulin * Deriving Complex Ligand Binding Formulas * Mathematical Modeling of Cooperative Interactions in Hemoglobin * Analysis of Interactions of Regulatory Protein TyrR with DNA * Parsing Free Energy of Drug-DNA Interactions * Use of Fluorescence as Thermodynamics Tool

Examining the physical basis of the structure of macromolecules—proteins, nucleic acids, and their complexes—using calorimetric techniques Many scientists working in biology are unfamiliar with the basics of thermodynamics and its role in determining molecular structures. Yet measuring the heat of structural change a molecule undergoes under various conditions yields information on the energies involved and, thus, on the physical bases of the considered structures. Microcalorimetry of Macromolecules offers protein scientists unique access to this important information. Divided into thirteen chapters, the book introduces readers to the basics of thermodynamics as it applies to calorimetry, the evolution of the calorimetric technique, as well as how calorimetric techniques are used in the thermodynamic studies of macromolecules, detailing instruments for measuring the heat effects of various processes. Also provided is general information on the structure of biological macromolecules, proteins, and nucleic acids, focusing on the key thermodynamic problems relating to their structure. The book covers: The use of supersensitive calorimetric instruments, including micro and nano-calorimeters for measuring the heat of isothermal reactions (Isothermal Titration Nano-Calorimeter), the heat capacities over a broad temperature range (Scanning Nano-Calorimeter), and pressure effects (Pressure Perturbation Nano-Calorimeter) Two of the simplest but key structural elements: the α and polyproline helices and their complexes, the α-helical coiled-coil, and the pyroline coiled-coils Complicated macromolecular formations, including small globular proteins, multidomain proteins and their complexes, and nucleic acids Numerous examples of measuring the ground state of protein energetics, as well as changes seen when proteins interact The book also reveals how intertwined structure and thermodynamics are in terms of a macromolecule's organization, mechanism of formation, the stabilization of its three-dimensional structure, and ultimately, its function. The first book to describe microcalorimetric technique in detail, enough for graduate students and research scientists to successfully plumb the structural mysteries of proteins and the double helix, Microcalorimetry of Macromolecules is an essential introduction to using a microcalorimeter in biological studies.

Over the last decade, high-sensitivity calorimetry has developed from a specialist method used mainly by dedicated experts to a major, commercially available tool in the arsenal directed at understanding molecular interactions and stability. Calorimeters have now become commonplace in bioscience laboratories. As a result, the number of those proficient in experimentation in this field has risen dramatically, as has the range of experiments to which these methods have been applied. Applications extend from studies in small molecule and solvent biophysics, through drug screening to whole cell assays. The technology has developed to include higher levels of sensitivity (and hence smaller sample size requirements) and a drive towards high-throughput technology, creating a very large user base in both academia and the pharmaceutical industry. This book is a fully revised and updated edition of the successful Biocalorimetry: Applications of Calorimetry in the Biological Sciences, published in 1998. Since then, there have been many advances in the instrumentation as well as in its applications and methodology. There are general chapters highlighting the usage of the isothermal titration calorimeter and the differential scanning calorimeter, more advanced chapters on specific applications and tutorials that cover the idiosyncrasies of experimental methods and data analysis. The book draws these together to create the definitive biological calorimetric text book. This book both explains the background to the method and describes novel, high-impact applications. It features works of interest to the experienced calorimetrist and the enthusiastic dilettante. The book should be of interest to all working in the field of biocalorimetry, from graduate students to researchers in academia and in industry.

Calorimetry, the latest volume in the Methods in Enzymology series continues the legacy of this premier serial with quality chapters authored by leaders in the field. Calorimetry is a highly technical experiment and it is easy for new practioners to get fooled into interpreting artifacts as real experimental results. This volume will guide readers to get the most out of their precious biological samples and includes topics on specific protocols for the types of studies being conducted as well as tips to improve the data collection. Most importantly, the chapters will also help to identify pitfalls that need to be avoided to ensure that the highest quality results are obtained. Contains timely contributions from recognized experts in this rapidly changing field Provides specific protocols and tips to improve data collection and ensure the highest quality results are obtained Covers research methods in calorimetry, and includes sections on topics such as differential scanning calorimetry of membrane and soluble proteins in detergents.