Macromolecular Crystallography Protocols Volume 1

Macromolecular Crystallography Protocols, now in two volumes, examines major developments that have occurred since publication of the acclaimed first edition nearly a decade ago. Volume 1 is composed of detailed protocols for the preparation and optimization of crystals. Volume 2 complements the first volume by addressing laboratory techniques for crystal handling and structural characterization. The volume concludes with a survey of available crystallographic software.

In the decade since the publication of the first edition of "Crystallographic Methods and Protocols", the field has seen several major developments that have both accelerated the pace of structure determination and made crystallography accessible to a broader range of investigators. "Volume I, Preparation and Crystallization of Macromolecules" is dedicated to the crystallization and ways to increase the odds of obtaining crystals in macromolecules, while "Volume 2, Structure Determination" covers both computational methods for characterizing crystals and solving structures.

In the decade since publication of the first edition this book, the field has seen several major developments. These developments have both accelerated the pace of structure determination and made crystallography accessible to a broader range of investigators. Volume I is dedicated to crystallization and ways to increase the odds of obtaining crystals in macromolecules. Volume 2 covers both computational methods for characterizing crystals and solving structures.

This volume provides methods for modern macromolecular crystallography, including all steps leading to crystal structure determination and analysis. 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, Protein Crystallography aims to ensure successful results in the further study of this vital field.

There has always been some tension between proponents of hypothesis-driven and discovery-driven research in the broad field of life sciences. Academic research has been primarily focused on hypothesis-driven research. However, the success of the human genome project, a discovery-driven research approach, has opened the door to adding other types of discovery-driven research to a continuum of research approaches. In contrast, drug discovery research in the pharmaceutical industry has embraced discovery-driven research for many years. A good example has been the discovery of active compounds from large chemical libraries, through screening campaigns. The success of the human genome project has also demonstrated the need for both academic researchers and industrial researchers to now understand the functions of genes and gene products. The cell is the basic unit of life and it has been at the cellular level where function can be demonstrated most cost-effectively and rapidly. High content screening (HCS) was developed by Cellomics Inc. in the mid-1990s to address the need for a platform that could be used in the discovery-driven research and development required to understand the functions of genes and gene products at the level of the cell.

Mitochondrial Genomics and Proteomics Protocols offers a broad collection of methods for studying the molecular biology, function, and features of mitochondria. In the past decade, mitochondrial research has elucidated the important influence of mitochondrial processes on integral cell processes such as apoptosis and cellular aging. This practical guide presents a wide spectrum of mitochondrial methods, each written by specialists with solid experience and intended for implementation by novice and expert researchers alike. Part I introduces major experimental model systems and discusses their specific advantages and limitations for functional analysis of mitochondria. The concise overview of general properties of mitochondrial systems is supplemented by detailed protocols for cultivation of model organisms. Parts II-VI comprise a robust collection of protocols for studying different molecular aspects of mitochondrial functions including: genetics and microbiology, biochemistry, physiology, dynamics and morphology, and functional genomics. Emphasis is placed on new and emerging topics in mitochondrial study, such as the examination of apoptotic effects, fusion and fission of mitochondria, and proteome and transcriptome analysis.

With the advent of high-throughput technologies following completion of the human genome project and similar projects, the number of genes of interest has expanded and the traditional methods for gene function analysis cannot achieve the throughput necessary for large-scale exploration. This book brings together a number of recently developed techniques for looking at gene function, including computational, biochemical and biological methods and protocols.

Macromolecular Crystallography is the study of macromolecules (proteins and nucleic acids) using X-ray crystallographic techniques in order to determine their molecular structure. The knowledge of accurate molecular structures is a pre-requisite for rational drug design, and for structure-based function studies to aid the development of effective therapeutic agents and drugs. The successful determination of the complete genome (genetic sequence) of several species (including humans) has recently directed scientific attention towards identifying the structure and function of the complete complement of proteins that make up that species; a new and rapidly growing field of study called 'structural genomics'. There are now several important and well-funded global initiatives in operation to identify all of the proteins of key model species. One of the main requirements for these initiatives is a high-throughput crystallization facility to speed-up the protein identification process. The extent to which these technologies have advanced, calls for an updated review of current crystallographic theory and practice. This practical reference book features the latest conventional and high-throughput methods, and includes contributions from a team of internationally recognized leaders and experts. It will be of relevance and use to graduate students, research scientists and professionals currently working in the field of conventional and high-throughput macromolecular crystallography.