Lipase and Phospholipase Protocols

About this book

A unique collection of step-by-step methods for the characterization, purification, and expression of mammalian lipases and phospholipases. The methods include the preparation of a variety of unique substrates for the determination of lipolytic activity, the isolation of purified lipase preparations, and the development of high expressing recombinant systems. There are also methods for the immunodectection and production of immunological reagents and the analysis of biochemical properties, such as subunit size, lipase kinetics, enzyme immobilization, and ligand interactions. The protocols covers more than a dozen specific enzymes, and provide well-established methods that can be adapted to the discovery of new lipase enzymes.

Contents

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 V'ronique 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 ?sterlund. 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 Andr' Bensadoun Jean Hsu and Barry Hughes. Purification of Rat Hepatic Lipase Essentially Free of Apolipoprotein E and Apolipoprotein B Andr' Bensadoun Barry Hughes Kristan Melford Jean Hsu and Dawn L. Braesaemle. High-Level Expression 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 V'ronique Lopez Alain de Caro Robert Verger and Fr'd'ric CarriSre. Modulation of the Expression Level of Human Acidic Lipases by Various Signal Peptides St'phane Canaan Liliane Dupuis Mireille RiviSre 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 V'ronique Lopez Mustapha Aoubala C'cile Daniel Robert Verger and Fr'd'ric CarriSre. 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 St'phane 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.

Customer Reviews