Lipidomics and bioactive lipids : lipids and cell signaling / edited by H. Alex Brown.

Format:

Book

Contributor:

Brown, H. Alex (Hugh Alex), 1960-2017.

Series:

Methods in enzymology ; v. 434.

Methods in enzymology ; v. 434

Language:

English

Subjects (All):

Lipids--Analysis.

Lipids.

Lipids--Spectra.

Mass spectrometry.

Physical Description:

xliv, 342 pages, 8 unnumbered pages of plates : illustrations (some color) ; 24 cm.

Place of Publication:

Amsterdam, Netherlands : Elsevier ; San Diego : Academic Press, [2007]

Contents:

Volumes in Series xix

1 Phospholipase A, Assays Using a Radiolabeled Substrate and Mass Spectrometry / Rei Morikawa, Masafumi Tsujimoto, Hiroyuki Arai, Junken Aoki 1

2 Types of PLA[subscript 1] 2

3 Conventional PLA[subscript 1] Assay Using Radiolabeled Substrates 3

4 Novel PLA[subscript 1] Assay Using ESI-MS 6

2 Real-Time Cell Assays of Phospholipase A[subscript 2]s Using Fluorogenic Phospholipids / Debasis Manna, Wonhwa Cho 15

2 Fluorogenic PLA[subscript 2] Substrates 18

3 Measuring Cellular sPLA[subscript 2] Activity Using PED6 and Red-PED6 23

4 Measuring Cellular cPLA[subscript 2 alpha] Activity Using DAPC 24

3 Analysis and Pharmacological Targeting of Phospholipase C [beta] Interactions with G Proteins / David M. Lehmann, Chujun Yuan, Alan V. Smrcka 29

4 Biochemical Analysis of Phospholipase D / H. Alex Brown, Lee G. Henage, Anita M. Preininger, Yun Xiang, John H. Exton 49

2 Assay of Recombinant PLD In Vitro 52

3 Regulated PLD1 Activity 59

4 Preparation of Activators of PLD1 60

5 Effects of Activators on PLD1 Activity 61

6 Synergy Between PLD1 Activators 62

7 Binding of PLD1 to Phospholipid Vesicles 63

8 Kinetic Parameters of PLD1 Catalytic Activity 65

9 Kinetic Analyses of Synergistic Responses 69

10 Phosphatidylinositol 4,5-Bisphosphate is an Essential PLD1 Activator 69

11 In ViVo PLD Assay Using Radioisotopes 74

12 In Vivo PLD Assay Using Deuterated 1-Butanol 74

13 Fluorescent In Vitro PLD Assay 77

14 Real-Time Diacylglycerol Lipase Assay 80

5 Measurement of Autotaxin/Lysophospholipase D Activity / Andrew J. Morris, Susan S. Smyth 89

2 Overview of Methods for Determination of Autotaxin/LysoPLD Activity 93

3 Expression of V5-Tagged Autotaxin/LysoPLD in HEK293 Cells 94

4 Measurement of Autotaxin/LysoPLD Activity Using Radiolabeled Substrates 95

5 Measurement of Autotaxin/LysoPLD Activity Using Fluorogenic Substrates 98

6 Platelet-Activating Factor / John S. Owen, Michael J. Thomas, Robert L. Wykle 105

2 Procedure 107

7 Quantitative Measurement of Phosphatidylinositol 3,4,5-trisphosphate / Herve Guillou, Len R. Stephens, Phillip T. Hawkins 117

2 Measuring Levels of Radioactively Labeled Phosphoinositides in Isolated Cells 120

3 Measuring Ptdlns(3,4,5)P[subscript 3] by Protein-Lipid Overlay 122

8 Measuring Phosphorylated Akt and Other Phosphoinositide 3-kinase-Regulated Phosphoproteins in Primary Lymphocytes / Amber C. Donahue, Michael G. Kharas, David A. Fruman 131

2 Choosing a Downstream Readout: General Considerations 134

3 Protocols for Detection of Pl[subscript 3]K-Regulated Phosphoproteins by Immunoblot 137

4 Protocols for Detection of Phosphoproteins by Flow Cytometry 142

9 Regulation of Phosphatidylinositol 4-Phosphate 5-kinase Activity by Partner Proteins / Yasunori Kanaho, Kazuhisa Nakayama, Michael A. Frohman, Takeaki Yokozeki 155

2 Protocols 158

10 Biochemical Analysis of Inositol Phosphate Kinases / James C. Otto, Sashidhar Mulugu, Peter C. Fridy, Shean-Tai Chiou, Blaine N. Armbruster, Anthony A. Ribeiro, John D. York 171

2 Experimental Methods 174

11 Analysis of Phosphoinositides and Their Aqueous Metabolites / Christopher P. Berrie, Cristiano Iurisci, Enza Piccolo, Renzo Bagnati, Daniela Corda 187

2 Cell Sample Extraction 191

3 Lipid Phase: TLC, HPLC Separation, and Desalting 197

4 Aqueous Phase: HPLC Separation, Desalting, and Ascintillant Extraction 203

5 Chemical Identification 212

6 ESI-MS/MS Identification 214

7 Standards 219

8 Final Considerations 226

12 Combination of C[subscript 17] Sphingoid Base Homologues Mass Spectrometry Analysis as a New Approach to Study Sphingolipid Metabolism / Stefka Spassieva, Jacek Bielawski, Viviana Anelli, Lina M. Obeid 233

2 Mass Spectrometry Analysis 235

3 Ceramide Synthase 236

4 In Vitro Ceramide Synthase Method 236

5 Sphingosine Kinase 237

6 In Vitro Sphingosine Kinase Method 238

7 In Cells Labeling with C[subscript 17] Sphingoid Base 239

13 Measurement of Mammalian Sphingosine-1-Phosphate Phosphohydrolase Activity In Vitro and In Vivo / Michael Maceyka, Sheldon Milstien, Sarah Spiegel 243

2 Principle 249

3 Measurement of SPP Activity in Cell Lysates 249

4 Measurement of SPP Activity in Live Cells 252

14 A Rapid and Sensitive Method to Measure Secretion of Sphingosine-1-Phosphate / Poulami Mitra, Shawn G. Payne, Sheldon Milstien, Sarah Spiegel 257

2 Measurement of S1P 259

3 Conclusions and Perspectives 262

15 Ceramide Kinase and Ceramide-1-Phosphate / Dayanjan S. Wijesinghe, Nadia F. Lamour, Antonio Gomez-Munoz, Charles E. Chalfant 265

2 Recombinant Expression and Kinetic Analysis of CERK 269

3 In Vitro Kinetic Analysis of CERK Activity Using Mixed Micellar Assays 272

4 Effective Delivery of C1P to Cells in Tissue Culture to Study Biological Effects 278

5 Analysis of Levels of Kinase-Derived C1P in Cells 281

6 Analysis of CERK Localization in Cells 284

7 Analysis of CERK Function by sIRNA-Mediated Manipulation of CERK Expression 286

8 Analysis of CERK mRNA Levels by Q-PCR 288

16 Measurement of Mammalian Diacylglycerol Kinase Activity In Vitro and in Cells / Richard M. Epand, Matthew K. Topham 293

2 In Vitro Assay of DGK 295

3 Measuring DGK Activity in Subcellular Compartments 300

4 Measuring DGK Activity in Cultured Cells 301

17 Lipid Phosphate Phosphatases from Saccharomyces cerevisiae / George M. Carman, Wen-I Wu 305

2 Preparation of Radiolabeled Substrates 307

3 Assay Methods 307

4 Growth of Yeast 308

5 Purification Procedure 308

6 Properties of DPP1- and LPP1-Encoded Lipid Phosphate Phosphatases 311.

Notes:

Includes bibliographical references and indexes.

ISBN:

9780123739650

0123739659

OCLC:

181350481