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Umfang: 1 online resource (387 p.)

ISBN: 9786611070731 , 9781281070739 , 1281070734 , 9780080554099 , 0080554091

Serie: Methods in enzymology ; v. 434

Inhalt: This volume in the well-established Methods in Enzymology series features methods for the study of lipids using mass spectrometry techniques. Articles in this volume cover topics such as Phospholipase A1 assays using a radio-labeled substrate and mass spectrometry; Real-time Cell Assays of Phospholipases A2 Using Fluorogenic Phospholipids; Analysis and Pharmacological Targeting of Phospholipase C â interactions with G proteins; Biochemical Analysis of Phospholipase D.; Measurement of Autotaxin/Lysophospholipase D Activity; Platelet-Activating Factor; Quantitative measurement of PtdIns(3

Anmerkung: Description based upon print version of record. , Cover; Contents; Contributors; Preface; Volumes in Series; Chapter 1: Phospholipase A1 Assays Using a Radiolabeled Substrate and Mass Spectrometry; 1. Introduction; 2. Types of PLA1; 3. Conventional PLA1 Assay Using Radiolabeled Substrates; 3.1. Materials; 3.2. Preparation of radiolabeled phospholipid substrates; 3.3. Assays; 4. Novel PLA1 Assay Using ESI-MS; 4.1. Materials; 4.2. Preparation of recombinant PLA1; 4.3. Phospholipase A1 assay; 4.4. PLA1 activity of intracellular PLA1s; 5. Perspective; Acknowledgments; References , Chapter 2: Real-Time Cell Assays of Phospholipase A2s Using Fluorogenic Phospholipids1. Introduction; 2. Fluorogenic PLA2 Substrates; 2.1. Substrate specificity of PLA2s; 2.2. Selection of fluorophores and design of fluorogenic PLA2 substrates; 2.3. Real-Time cellular PLA2 assay using fluorogenic phospholipids; 3. Measuring Cellular sPLA2 Activity Using PED6 and Red-PED6; 3.1. Materials; 3.2. Labeling of cell membranes with PED6 and monitoring cellular sPLA2 activities; 4. Measuring Cellular cPLA2alpha Activity Using DAPC; 4.1. Synthesis of DAPC , 4.2. Labeling of cell membranes with DAPC and monitoring cellular cPLA2alpha activityReferences; Chapter 3: Analysis and Pharmacological Targeting of Phospholipase C beta Interactions with G Proteins; 1. Introduction; 2. Methods; 2.1. Protein expression and purification; 2.2. PLCbeta assay; 2.3. PLC assay optimization; 2.4. Application of the PLC assay to evaluate peptide/small molecule modulation of Gbetagamma-dependent PLC activity; 2.5. Evaluation of Gbetagamma-PLC binding; 3. Concluding Remarks; Acknowledgment; References; Chapter 4: Biochemical Analysis of Phospholipase D , 1. Introduction2. Assay of Recombinant PLD In Vitro; 2.1. Expression of recombinant PLD1; 2.2. Purification of PLD1; 2.3. Chromatography (6His); 2.4. Size-exclusion chromatography (Superdex 200); 2.5. Anion-exchange chromatography (Q-Sepharose); 2.6. Characteristics of purified, full-length PLD1; 2.7. Purification of N-terminally truncated PLD1; 2.8. Assay of phospholipase D activity in vitro; 3. Regulated PLD1 Activity; 4. Preparation of Activators of PLD1; 4.1. Protein kinase Calpha (PKCalpha); 4.2. Myristoyl-ADP ribosylation factor 1 , 4.3. Purification of geranylgeranylated RhoA, Rac1, and Cdc425. Effects of Activators on PLD1 Activity; 6. Synergy between PLD1 Activators; 7. Binding of PLD1 to Phospholipid Vesicles; 8. Kinetic Parameters of PLD1 Catalytic Activity; 9. Kinetic Analyses of Synergistic Responses; 10. Phosphatidylinositol 4,5-Bisphosphate is an Essential PLD1 Activator; 11. In Vivo PLD Assay Using Radioisotopes; 12. In Vivo PLD Assay Using Deuterated 1-Butanol; 12.1. Materials and methods; 12.2. Application; 13. Fluorescent In Vitro PLD Assay; 13.1. Preparation of lipid substrates; 13.2. Phospholipase D assay , 14. Real-Time Diacylglycerol Lipase Assay , English

Weitere Ausg.: ISBN 9780123739650

Weitere Ausg.: ISBN 0123739659

Sprache: Englisch

UID:

Umfang: Online-Ressource (342 S.)

Ausgabe: Online-Ausg. 2009 Electronic reproduction; Available via World Wide Web

ISBN: 9780123739650

Serie: Methods in enzymology; 434

Inhalt: This volume in the well-established Methods in Enzymologyseries features methods for the study of lipids using mass spectrometry techniques. Articles in this volume cover topics such as Phospholipase A1 assays using a radio-labeled substrate and mass spectrometry, Real-time Cell Assays of Phospholipases A2 Using Fluorogenic Phospholipids, Analysis and Pharmacological Targeting of Phospholipase C ? interactions with G proteins, Biochemical Analysis of Phospholipase D., Measurement of Autotaxin/Lysophospholipase D Activity, Platelet-Activating Factor, Quantitative measurement of PtdIns(3,4,5)P3, Measuring Phosphorylated Akt And Other Phosphoinositide 3-Kinase-Regulated Phosphoproteins In Primary Lymphocytes, Regulation of Phosphatidylinositol 4-Phosphate 5-Kinase activity by partner proteins, Biochemical Analysis of Inositol Phosphate Kinases, Analysis of the phosphoinositides and their aqueous metabolites, Combination of C17-sphingoid base homologues and mass spectrometry analysis as a new approach to study sphingolipid metabolism, Measurement of mammalian sphingosine-1-phosphate phosphohydrolase activity in vitro and in vivo, A rapid and sensitive method to measure secretion of sphingosine-1-phosphate, Ceramide Kinase and Ceramide-1-Phosphate, Measurement of Mammalian Diacylglycerol Kinase Activity in vitro and in Cells, Lipid Phosphate Phosphatases from Saccharomyces cerevisiae.

Anmerkung: Includes bibliographical references and index , Electronic reproduction; Available via World Wide Web

Weitere Ausg.: Erscheint auch als Druck-Ausgabe Lipidomics and bioactive lipids: Lipids and cell signaling Amsterdam [u.a.] : Elsevier, Acad. Press, 2007 ISBN 9780123739650

Sprache: Englisch

Fachgebiete: Biologie

Schlagwort(e): Lipidstoffwechsel ; Zelle ; Signaltransduktion

URL: Volltext

URL: Inhaltsverzeichnis

UID:

Umfang: 1 Online-Ressource

ISBN: 9780123739650 , 0123739659

Serie: Methods in enzymology 434

Sprache: Englisch

Fachgebiete: Biologie

Schlagwort(e): Lipide ; Signaltransduktion ; Labortechnik ; Aufsatzsammlung

URL: Volltext