scholarly journals Synthesis and Evaluation of Fluorogenic Substrates for Phospholipase D and Phospholipase C

2006 ◽  
Vol 8 (12) ◽  
pp. 2575-2578 ◽  
Author(s):  
Tyler M. Rose ◽  
Glenn D. Prestwich
Keyword(s):  
Phospholipase C ◽  
Phospholipase D ◽  
Fluorogenic Substrates

scholarly journals Targeting of Protein Kinase C-ϵ during Fcγ Receptor-dependent Phagocytosis Requires the ϵC1B Domain and Phospholipase C-γ1

2006 ◽  
Vol 17 (2) ◽  
pp. 799-813 ◽  
Author(s):  
Keylon L. Cheeseman ◽  
Takehiko Ueyama ◽  
Tanya M. Michaud ◽  
Kaori Kashiwagi ◽  
Demin Wang ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phospholipase C ◽  
Phospholipase D ◽  
Fluorescent Protein ◽  
Wild Type ◽  
Fcγ Receptor ◽  
Kinase C ◽  
Green Fluorescent

Protein kinase C-ϵ (PKC-ϵ) translocates to phagosomes and promotes uptake of IgG-opsonized targets. To identify the regions responsible for this concentration, green fluorescent protein (GFP)-protein kinase C-ϵ mutants were tracked during phagocytosis and in response to exogenous lipids. Deletion of the diacylglycerol (DAG)-binding ϵC1 and ϵC1B domains, or the ϵC1B point mutant ϵC259G, decreased accumulation at phagosomes and membrane translocation in response to exogenous DAG. Quantitation of GFP revealed that ϵC259G, ϵC1, and ϵC1B accumulation at phagosomes was significantly less than that of intact PKC-ϵ. Also, the DAG antagonist 1-hexadecyl-2-acetyl glycerol (EI-150) blocked PKC-ϵ translocation. Thus, DAG binding to ϵC1B is necessary for PKC-ϵ translocation. The role of phospholipase D (PLD), phosphatidylinositol-specific phospholipase C (PI-PLC)-γ1, and PI-PLC-γ2 in PKC-ϵ accumulation was assessed. Although GFP-PLD2 localized to phagosomes and enhanced phagocytosis, PLD inhibition did not alter target ingestion or PKC-ϵ localization. In contrast, the PI-PLC inhibitor U73122 decreased both phagocytosis and PKC-ϵ accumulation. Although expression of PI-PLC-γ2 is higher than that of PI-PLC-γ1, PI-PLC-γ1 but not PI-PLC-γ2 consistently concentrated at phagosomes. Macrophages from PI-PLC-γ2-/-mice were similar to wild-type macrophages in their rate and extent of phagocytosis, their accumulation of PKC-ϵ at the phagosome, and their sensitivity to U73122. This implicates PI-PLC-γ1 as the enzyme that supports PKC-ϵ localization and phagocytosis. That PI-PLC-γ1 was transiently tyrosine phosphorylated in nascent phagosomes is consistent with this conclusion. Together, these results support a model in which PI-PLC-γ1 provides DAG that binds to ϵC1B, facilitating PKC-ϵ localization to phagosomes for efficient IgG-mediated phagocytosis.

Phosphatidic acid production in chitosan-elicited tomato cells, via both phospholipase D and phospholipase C/diacylglycerol kinase, requires nitric oxide

2011 ◽  
Vol 168 (6) ◽  
pp. 534-539 ◽  
Author(s):  
Nicolás Raho ◽  
Leonor Ramirez ◽  
M. Luciana Lanteri ◽  
Gabriela Gonorazky ◽  
Lorenzo Lamattina ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Phosphatidic Acid ◽  
Phospholipase C ◽  
Phospholipase D ◽  
Acid Production ◽  
Diacylglycerol Kinase

scholarly journals Allosteric Interactions within Subsites of a Monomeric Enzyme: Kinetics of Fluorogenic Substrates of PI-Specific Phospholipase C

2003 ◽  
Vol 84 (5) ◽  
pp. 3264-3275 ◽  
Author(s):  
G. Bruce Birrell ◽  
Tatiana O. Zaikova ◽  
Aleksey V. Rukavishnikov ◽  
John F.W. Keana ◽  
O. Hayes Griffith
Keyword(s):  
Enzyme Kinetics ◽  
Phospholipase C ◽  
Fluorogenic Substrates ◽  
Allosteric Interactions ◽  
Kinetics Of
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