Presence of a phospholipase D (PLD) distinct from PLD1 or PLD2 in human neutrophils: immunobiochemical characterization and initial purification

• Published in: Biochimica et biophysica acta Vol. 1530; no. 1; pp. 97 - 110
• Main Authors: Horn, Jeffrey M, Lehman, Jason A, Alter, Gerald, Horwitz, Joel, Gomez-Cambronero, Julian
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.01.2001
• Subjects: Adenosine Triphosphate; Cations, Divalent; Chromatography, Ion Exchange; Cloning, Molecular; Detergents; Dithiothreitol; Enzyme characterization; Enzyme Inhibitors - pharmacology; Guanosine 5'-O-(3-Thiotriphosphate); Humans; Hydrogen Peroxide; Hydrogen-Ion Concentration; Kinetics; Neutrophil; Neutrophils - enzymology; Oleic Acid - pharmacology; Phospholipase D; Phospholipase D - analysis; Phospholipase D - genetics; Phospholipase D - isolation & purification; Phospholipase D - metabolism; Phosphotyrosine; Signal transduction; PC, phosphatidylcholine; PLC, phospholipase C; PC8, 1,2-dioctanoyl- sn-glycero-3-phosphocholine; PKC, protein kinase C; Phosphotyrosine; DAG, diacylglycerol; PIP 2, L-α-phosphatidyl- D- myo-inositol-4,5- bis-phosphate; Phospholipase D; Enzyme characterization; PLA 2, phospholipase A 2; Signal transduction; PY, phosphotyrosine; PLD, phospholipase D; Neutrophil; PA, phosphatidic acid; ARF, ADP-ribosylating factor

Utilizing the transphosphatidylation reaction catalyzed by phospholipase D (PLD) in the presence of a primary alcohol and the short-chain phospholipid PC8, we have characterized the enzyme from human neutrophils. A pH optimum of 7.8–8.0 was determined. PIP 2, EDTA/EGTA, and ATP were found to enhance...