David Morgan brings together in Polyamine Protocols a unique collection of hands-on enzyme assay techniques for the study of polyamines and their biological functions. The techniques range from assay methods for enzymes of polyamine biosynthesis and catabolism to measurements of polyamines, polyamine transport, and polyamine effects on cell growth. All methods are presented by active researchers who have developed them to a high degree of perfection in their own laboratories, and include clear, step-by-step instructions with numerous hints and tips to ensure readily reproducible results.

Polyamine Protocols is the only compilation of methods currently available for use in studies involving polyamines. The collection offers both novice and experienced researchers across many fields-including cell and molecular biology, pharmacology, oncology, physiology, laboratory medicine, and clinical pathology-all the tools needed to elucidate the biological roles played by these ubiquitous substances.

Klappentext

Putrescine and spermidine are ubiquitous in living organisms. Spermine, third of the three most commonly occurring natural polyamines, is probably present in all eukaryotes but is rare (or nonexistent) in prokaryotes. Polyamine residues are constituents of many compounds found in plants and insects. Putrescine, spermidine, or spermine-containing alkaloids are found in many plants, nonproteinaceous spider and wasp toxins contain polyamine residues, and glutathionyl spermidine conjugates have been found in some pathogenic microorganisms. In most cells polyamines are the products of a highly regulated bios- thetic pathway. It is not clear whether the elaborate regulation of polyamine synthesis is a consequence of their essential role(s) in cellular differentiation and development, or part of a defense mechanism to prevent overaccumulation of compounds that are toxic in excess. In addition to their biosynthetic capa­ bility, many cells also possess transport systems for polyamines that respond to intracellular polyamine levels, and other stimuli, and are regulated by mecha­ nisms that are at present incompletely defined. Two routes of polyamine catabolism have been identified in mammalian cells, a biodegradative route and a recycling pathway. The relative impor­ tance of these pathways and their overall regulation is only partially resolved. What is clear is the widespread occurrence of a variety of polyamine-oxid- ing enzymes in animals, plants, bacteria, and fungi. Polyamine catabolism, by whichever route, results in the formation of aminoaldehydes as intermediates.

Inhalt
Polyamines: An Introduction.- Assay Methods for Enzymes of Polyamine Biosynthesis.- Determination of Ornithine Decarboxylase Activity Using [3H]Omithine.- Assay of Mammalian Ornithine Decarboxylase Activity Using [14C]Ornithine.- Assay of Mammalian S-Adenosylmethionine Decarboxylase Activity.- Assay of Spermidine and Spermine Synthases.- Measurement of Spermidine/Spermine N 1-Acetyltransferase Activity.- Assay of Spermidine N 8-Acetyltransferase.- Assay Methods for Enzymes of Polyamine Catabolism.- Assay of N 8-Acetylspermidine Deacetylase.- Hydrogen Peroxide Assay for Amine Oxidase Activity.- Radiochemical Assay of Diamine Oxidase.- Radiochemical Estimation of Polyamine Oxidase.- Measurement of Aldehyde Formation as an Assay for Polyamine Oxidase Activity.- Measurement of Polyamines.- Determination of Polyamines as Their Benzoylated Derivatives by HPLC.- A Dansyl Chloride-HPLC Method for the Determination of Polyamines.- The Determination of Polyamines and Amino Acids by a Fluorescamine-HPLC Method.- Thin-Layer Chromatographic Method for Assaying Polyamines.- Measurement of Polyamine Transport.- Measurement of Polyamine Transport: Adherent Cells.- Measurement of Polyamine Transport: Cells in Suspension.- Measurement of Polyamine Efflux from Cells in Culture.- Measurement of Polyamine Effects on Cell Growth.- Leucine Incorporation and Thymidine Incorporation.- Tetrazolium (MTT) Assay for Cellular Viability and Activity.