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Conditionally Toxic Proteins
Human health depends upon access to high-quality proteins for our diet and pharmaceutical use. This book deals with the nature of toxicity as it applies to proteins in food and drugs. Many proteins, such as glutens and allergens, are valuable food sources but toxic for sensitized individuals. Even proteins produced in the human body can become toxic through mutation or aggregation. For example, Alzheimer's and Huntington's disease are characterized by plaques in the brain of insoluble protein aggregations. Paradoxically, even toxins produced by the most pathogenic microorganisms, such as Botox, have found use in the clinic and industry. This book discusses how many proteins, including interf...
Mycotoxin Protocols
Mycotoxins produced by molds are common contaminants of many important crops, including wheat, corn, rice, and peanuts. Some mycotoxins are found in fruits and vegetables. These contaminants have a broad range of toxic effects, including carcinogenicity, neurotoxicity, and reproductive and developmental toxicity. The occurrence of mycotoxins in foods is an unavoidable worldwide problem. About 80 countries have imposed regulatory limits to minimize human and animal exposure to mycotoxins. Regulatory limits, including international standards, have tremendous economic impact and must be developed using science-based risk assessments. The purpose of Mycotoxin Protocols is to provide the scientif...
Steroid Receptor Methods
A distinguished team of principal investigators and their associates describe in step-by-step detail a cross-section of the latest research techniques available for studying the endocrine system. As a basis for sophisticated biochemical analysis of receptor properties, the contributors provide methods for the production and purification of a variety of receptors, including progesterone, glucocorticoid, and androgen. Other protocols allow the reader to experiment with DNA binding characteristics, hormone binding assays, and the use of combinatorial chemistry for drug discovery. A series of novel methods utilizing the latest advances in immunochemistry, yeast two-hybrid screening, and fluorescence are included for the detection and analysis of a variety of cellular proteins that influence steroid receptor effectiveness.
Affinity Chromatography
Affinity chromatography, with its exquisite specificity, is based upon molecular recognition. It is a powerful tool for the purification of biomolecules. In recent years, numerous new applications and modified techniques have been derived from gro- specific interactions and biological recognition principles. An up-to-date review of the past, current, and future applications of affinity chromatography has been presented in the introductory chapter by Meir Wilchek and Irwin Chaiken. Though many of these new applications and techniques are well documented in the literature, it is often difficult to find methods that are written with the intent of helping new practitioners of affinity chromatogr...
Protein Structure, Stability, and Folding
In Protein Structure, Stability, and Folding, Kenneth P. Murphy and a panel of internationally recognized investigators describe some of the newest experimental and theoretical methods for investigating these critical events and processes. Among the techniques discussed are the many methods for calculating many of protein stability and dynamics from knowledge of the structure, and for performing molecular dynamics simulations of protein unfolding. New experimental approaches presented include the use of co-solvents, novel applications of hydrogen exchange techniques, temperature-jump methods for looking at folding events, and new strategies for mutagenesis experiments. Unique in its powerful combination of theory and practice, Protein Structure, Stability, and Folding offers protein and biophysical chemists the means to gain a more comprehensive understanding of some of this complex area by detailing many of the major techniques in use today.
Connexin Methods and Protocols
Direct cell–cell communication is a common property of multicellular organisms that is achieved through membrane channels which are organized in gap junctions. The protein subunits of these intercellular channels, the connexins, form a multigene family that has been investigated in great detail in recent years. It has now become clear that, in different tissues, connexins speak several languages that control specific cellular functions. This progress has been made possible by the availability of new molecular tools and the improvement of basic techniques for the study of membrane channels, as well as by the use of genetic approaches to study protein function in vivo. More important, connex...
Two-Hybrid Systems
The yeast two-hybrid system is one of the most widely used and productive techniques available for investigating the macromolecular interactions that affect virtually all biological processes. In Two-Hybrid Systems: Methods and Protocols, Paul N. MacDonald has assembled a collection of these powerful molecular tools for examining and characterizing protein-protein, protein-DNA, and protein-RNA interactions. The techniques range from the most basic (introducing plasmids into yeasts, interaction assays, and recovering the plasmids from yeast) to the most advanced alternative strategies (involving one-hybrid, split two-hybrid, three-hybrid, membrane recruitment systems, and mammalian systems). ...
Proteoglycan Protocols
Proteoglycans are some of the most elaborate macromolecules of mammalian and lower organisms. The covalent attachment of at least five types of glycosami- glycan side chains to more than forty individual protein cores makes these molecules quite complex and endows them with a multitude of biological functions. Proteoglycan Protocols offers a comprehensive and up-to-date collection of prepa- tive and analytical methods for the in-depth analysis of proteoglycans. Featuring st- by-step detailed protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, to g...
Extracellular Matrix Protocols
It is now widely accepted that much of the dynamic function of cells and tissues is regulated from outside the cell by the extracellular matrix. In ad- tion to its conventional role in providing a scaffold for building tissues, the extracellular matrix acts as a directional highway for cellular movement and provides instructional information for promoting survival, proliferation, and differentiation. Indeed, the extracellular matrix is beginning to take a starring role in the choreography of cell and tissue function. The diverse roles of the extracellular matrix are reflected in its highly complicated structure, consisting of an ever increasing number of components. Yet the mechanisms of ext...
Gene Knockout Protocols
As the major task of sequencing the human genome is near completion and full complement of human genes are catalogued, attention will be focused on the ultimate goal: to understand the normal biological functions of these genes, and how alterations lead to disease states. In this task there is a severe limitation in working with human material, but the mouse has been adopted as the favored animal model because of the available genetic resources and the highly conserved gene conservation linkage organization. In just of ten years since the first gene-targeting experiments were p- formed in embryonic stem (ES) cells and mutations transmitted through the mouse germline, more than a thousand mou...
