Lipase and Phospholipase Protocols

ISBN-13: 9781617370694 / Angielski / Miękka / 2010 / 362 str.

Mark Doolittle

Opis

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.

Recenzje

"...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

Spis treści

Part I. Lipase Activity, Purification, and Expression. Phospholipase A2 and Phosphatidylinositol- Specific Phospholipase C Assays by HPLC and TLC with Fluorescent Substrate, H. Stewart Hendrickson. Fluorometric Phospholipase Assays Based on Polymerized Liposome Substrates, Wonhwa Cho, Shih-Kwang Wu, Edward Yoon, and Lenka Lichtenbergova. Triglyceride Lipase Assays Based on a Novel Fluorogenic Alkyldiacyl Glycerol Substrate, Albin Hermetter. Purification and Assay of Mammalian Group I and Group IIa Secretory Phospholipase A2, Wonhwa Cho, Sang Kyou Han, Byung-In Lee, Yana Snitko, and Rajiv Dua. Determination of Plasmalogen-Selective Phospholipase A2 Activity by Radiochemical and Fluorometric Assay Procedures, Akhlaq A. Farooqui, Hsiu-Chiung Yang, Yutaka Hirashima, and Lloyd A. Horrocks. Human Plasma Platelet-Activating Factor Acetylhydrolase, Diana M. Stafforini and Larry W. Tjoelker. Assays for Pancreatic Triglyceride Lipase and Colipase, Mark E. Lowe. Bile Salt-Activated Lipase, Chi-Sun Wang, Azar Dashti, and Deborah Downs. Determining Lipoprotein Lipase and Hepatic Lipase Activity Using Radiolabeled Substrates, Véronique Briquet-Laugier, Osnat Ben-Zeev, and Mark H. Doolittle. Lysosomal Acid Lipase: Assay and Purification, Martin Merkel, Anne-Christine Tilkorn, Heiner Greten, and Detlev Ameis. Hormone-Sensitive Lipase and Neutral Cholesteryl Ester Lipase, Cecilia Holm and Torben Østerlund. Lecithin-Cholesterol Acyltransferase: Assay of Cholesterol Esterification and Phospholipase A2 Activities, John S. Parks, Abraham K. Gebre, and James W. Furbee. Purification of Lipases and Phospholipases by Heparin-Sepharose Chromatography, Akhlaq A. Farooqui and Lloyd A. Horrocks. Large-Scale Lipoprotein Lipase Purification from Adipose Tissue, André Bensadoun, Jean Hsu, and Barry Hughes. Purification of Rat Hepatic Lipase Essentially Free of Apolipoprotein E and Apolipoprotein B, André Bensadoun, Barry Hughes, Kristan Melford, Jean Hsu, and Dawn L. Braesaemle. High-LevelExpression and Purification of Human Hepatic Lipase from Mammalian Cells, John S. Hill. High-Level Baculoviral Expression of Hormone-Sensitive Lipase, Cecilia Holm and Juan Antonio Contreras. High-Level Baculoviral Expression of Lysosomal Acid Lipase, Anne-Christine Tilkorn, Martin Merkel, Heiner Greten, and Detlev Ameis. One-Step Purification and Biochemical Characterization of Recombinant Pancreatic Lipases Expressed in Insect Cells, Sofiane Bezzine, Francine Ferrato, Véronique Lopez, Alain de Caro, Robert Verger, and Frédéric Carrière. Modulation of the Expression Level of Human Acidic Lipases by Various Signal Peptides, Stéphane Canaan, Liliane Dupuis, Mireille Rivière, Robert Verger, and Catherine Wicker-Planquart. Part II. Biochemical Characterization. Immunodetection of Lipoprotein Lipase: Antibody Production, Immunoprecipitation, and Western Blotting Techniques, Mark H. Doolittle and Osnat Ben-Zeev. Immunological Characterization of Digestive Lipases, Alain De Caro, Sofiane Bezzine, Véronique Lopez, Mustapha Aoubala, Cécile Daniel, Robert Verger, and Frédéric Carrière. Determining Lipase Subunit Structure by Sucrose Gradient Centrifugation, Osnat Ben-Zeev and Mark H. Doolittle. Detecting Ligands Interacting with Lipoprotein Lipase, Sivaram Pillarisetti. Monolayer Techniques for Studying Lipase Kinetics, Stéphane Ransac, Margarita Ivanova, Ivan Panaiotov, and Robert Verger. Efficient Immobilization of Phospholipase A2, Wonhwa Cho and Zhen Shen. Part III. Molecular Genetic Characterization. An Introduction to Internet Resources for the Molecular and Genetic Analysis of the Lipases, Karen Reue. Techniques for the Measurement of Lipoprotein Lipase Messenger RNA, Gouri Ranganathan and Philip A. Kern. In Vitro Transcription and Translation of Lipoprotein Lipase, Gouri Ranganathan and Philip A. Kern. Induced Lipase Mutations in the Mouse, Clay F. Semenkovich. Index.

Blurb

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.