Lipase and Phospholipase Protocols

Beschreibung

Lipase and Phospholipase Protocols is an important new compilation of cutting-edge techniques and strategies for the characterization, purification, and expression of mammalian lipases and phospholipases. The methods include the preparation of a variety of radiolabeled and fluorescent substrates for the determination of lipolytic activity, proven strategies for the expression and purification of recombinant lipases and phospholipases, and the characterization of lipases by such techniques as immunodetection, subunit size determination, lipase kinetics, and ligand interactions. In addition, there are methods for the molecular and genetic analysis of the lipases, the quantification of mRNA levels, and in vitro-coupled transcription/translation. The protocols cover more than a dozen specific enzymes, and many can be applied directly to all lipases and phospholipases.

Lipase and Phospholipase Protocols constitutes a highly useful single-volume handbook of basic procedures and strategies to investigate this unique class of enzymes. Its step-by-step format works perfectly at the laboratory bench or by the computer, providing practical information and examples that will lead to successful design and execution of specific techniques. It not only furnishes a technical framework for the investigation of known lipases and phospholipases, but provides well-established methods readily daptable to the discovery of new lipase enzymes.




The lipases and phospholipases represent a diverse group of enzymes that are expressed in animals, plants, fungi, and bacteria. Their ubiquitous distribution among all species is a testament to the essential roles played by these enzymes in lipid storage, mobilization and metabolism, membrane homeostasis and remodeling, endocrine and immune functions, and signal tra- duction. In humans, lipases and phospholipases are also thought to contribute to complex diseases, such as atherosclerosis, obesity, arthritis, and cancer, as well as to single gene defects, such as Wolman's disease and Type I hyperlipoproteinemia. Enzymatically, the lipases are unique, since they hydrolyze substrates that are either insoluble, or only partly soluble, in aq- ous solvents; thus, enzyme catalysis takes place at a lipid-water interface. The interface comprises at least two, and often more, discrete bulk and s- face phases, in which the enzyme, substrate, and products oflipolysis disperse among these phases based on their physical properties. Furthermore, the d- tribution of these components changes continuously as lipolysis proceeds. Thus, the lipases and phospholipases are fundamentally different from any other enzyme because of the physical complexity of the environment in which catalysis occurs.

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Mark Doolittle

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"...The book can be recommended to those scientists who are deeply engaged in lipid chemistry and analysis as well as enzymology. It provides a detailed listing of literature and other sources. Thus it can be successfully used as an introduction to new research fields and experimental techniques..."-Nahrung

"In view of the rapid increase in the number of new lipases and phospholipases that have been recently identified, the focus of this book is timely...In this volume, modern, up-to-date protocols with sufficient detail for experienced or peripheral investigators in this area are provided, and should prove to be a valuable resource."-The Quarterly Review of Biology
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