Purification and characterization of <i>sn</i>-1-stearoyl-2-arachidonoylglycerol kinase from pig testes

Hodgkin, Matthew N.; Gardner, Sandra D.; Rose, Sally; Paterson, Andrew J.; Martin, Ashley; Wakelam, Michael J.O.

doi:10.1042/bj3220529

Cited by 8 publications

(3 citation statements)

References 41 publications

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“…We could not present direct evidence that arachidonic acid released by PLA # after stimulation with SPC does indeed derive from phosphatidic acid generated by DAG kinase. However, we showed that the levels of polyunsaturated DAG and phosphatidic acid species increased after SPC stimulation, as reported by others [25,46], and this suggests that it is the polyunsaturated DAG that is converted predominantly to phosphatidic acid by some isoforms of DAG kinase [44,47]. Since the pertussis toxin-sensitive phosphatidic acid production by SPC in CFNP9o − cells was revealed only in the presence of AACOCF $ , this suggests that arachidonic acid might be released from DAG kinase-generated phosphatidic acid, producing lysophosphatidic acid, which seems therefore to be the intracellular messenger able to release Ca# + from internal stores.…”

Section: Discussionsupporting

confidence: 88%

Intracellular calcium mobilization and phospholipid degradation in sphingosylphosphorylcholine-stimulated human airway epithelial cells

Orlati

Porcelli

Hrelia

et al. 1998

Biochemical Journal

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Extracellular sphingosylphosphorylcholine (SPC) caused a remarkable elevation in the intracellular Ca2+ concentration ([Ca2+]i) in immortalized human airway epithelial cells (CFNP9o-). An increase in total inositol phosphates formation was determined; however, the dose responses for [Ca2+]i elevation and inositol phosphates production were slightly different and, furthermore, PMA and pertussis toxin almost completely inhibited [Ca2+]i mobilization by SPC, whereas inositol phosphates production was only partially reduced. The possible direct interaction of SPC with Ca2+ channels of intracellular stores was determined by experiments with permeabilized cells, where SPC failed to evoke Ca2+ release, whereas lysophosphatidic acid was shown to be effective. The level of phosphatidic acid was increased by SPC only in the presence of AACOCF3, a specific inhibitor of phospholipase A2 (PLA2) and blocked by both pertussis toxin and R59022, an inhibitor of diacylglycerol kinase. R59022 enhanced diacylglycerol production by SPC and also significantly reduced [Ca2+]i mobilization. Only polyunsaturated diacylglycerol and phosphatidic acid were generated by SPC. Lastly, SPC caused stimulation of arachidonic acid release, indicating the involvement of PLA2. Taken together, these data suggest that, after SPC stimulation, phospholipase C-derived diacylglycerol is phosphorylated by a diacylglycerol kinase to phosphatidic acid, which is further hydrolysed by PLA2 activity to arachidonic and lysophosphatidic acids. We propose that lysophosphatidic acid might be the intracellular messenger able to release Ca2+ from internal stores.

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Section: Discussionsupporting

confidence: 88%

Intracellular calcium mobilization and phospholipid degradation in sphingosylphosphorylcholine-stimulated human airway epithelial cells

Orlati

Porcelli

Hrelia

et al. 1998

Biochemical Journal

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“…In PAE cells there is a large amount of polyunsaturated DAG (over 25% of total DAG) [23]. In vitro studies have shown that the DGKϵ isoform is the only DGK isoform which shows specificity for a DAG containing a polyunsaturated fatty acid [25–27]. This specificity has been the subject of a very recent in vivo study confirming the selective removal of solely polyunsaturated DAG species by DGKϵ in stably DGKϵ‐transfected PAE cells [28].…”

Section: Resultsmentioning

confidence: 91%

Type Iα phosphatidylinositol 4‐phosphate 5‐kinase is a putative target for increased intracellular phosphatidic acid

2000

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Despite the fact that phosphatidic acid (PtdOH) has been implicated as a lipid second messenger for nearly a decade, its intracellular targets have remained unclear. We sought to investigate how an increase in the level of PtdOH could modulate phosphatidylinositol 4-phosphate 5-kinase (PIPkin), an enzyme involved in phosphatidylinositol 4,5-bisphosphate synthesis. Transfection of porcine aortic endothelial (PAE) cells with haemagglutinin (HA)-tagged type IK K PIPkin followed by immunofluorescence confocal microscopy revealed the enzyme to be localised to the plasma membrane. When the transfected PAE cells were stimulated with lyso-PtdOH, increased PIPkin activity was found to be associated with HA immunoprecipitates in an in vitro assay. This PIPkin activation was found to be greatly reduced by prior treatment of the cells with 1-butanol, thereby implicating phospholipase D (PLD) as the in vivo generator of PtdOH. In order to determine if the PtdOHdependent activation of type IK K PIPkin was dictated by a specific molecular composition of PtdOH, the wild type murine and porcine K K isoforms of diacylglycerol kinase (DGK) were individually co-transfected along with type IK K PIPkin. Under these conditions an increase in type IK K PIPkin lipid kinase activity was found in HA immunoprecipitates in an in vitro assay. No increases in lipid kinase activity were observed when type IK K PIPkin was co-transfected with either the human DGKO O isoform or a kinase-dead mutant of the murine DGKK K isoform. These results provide the first direct evidence for the unification of the production of saturated/monounsaturated PtdOH (through two different routes, PLD and DGK) and the in vivo activation of type IK K PIPkin by this lipid second messenger. ß

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“…Another example for highly specific interactions of soluble protein with lipid is the specific recognition of a single molecular diacylglycerol (DAG) species by a DAG-kinase in pig testes [55]. Its human homologue, DGK, was shown to be highly selective for arachidonate-containing species of DAG, providing a mechanism by which the cell can inactivate the lipid branch of the phosphoinositide-signalling pathway.…”

Section: Highly Specific Protein-lipid Interactions Of Soluble Proteinsmentioning

confidence: 99%

Determinants of specificity at the protein–lipid interface in membranes

et al. 2010

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a b s t r a c tThe complexity of pro-and eukaryotic lipidomes is increasingly appreciated mainly owing to the advance of mass spectrometric methods. Biophysical approaches have revealed that the large number of lipid classes and molecular species detected have implications for the self-organizing potential of biological membranes, resulting in the formation of lateral heterogeneous phases. How membrane proteins are able to adapt specifically to their surrounding heterogeneous matrix, and whether this environment affects protein targeting and function, is therefore a matter of particular interest. Here, we review specific protein-lipid interactions, focusing on the molecular mechanisms that determine specificity at the protein-lipid interface, and on membrane proteins that require lipids as cofactors for their architecture and function.

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Purification and characterization of sn-1-stearoyl-2-arachidonoylglycerol kinase from pig testes

Cited by 8 publications

References 41 publications

Intracellular calcium mobilization and phospholipid degradation in sphingosylphosphorylcholine-stimulated human airway epithelial cells

Intracellular calcium mobilization and phospholipid degradation in sphingosylphosphorylcholine-stimulated human airway epithelial cells

Type Iα phosphatidylinositol 4‐phosphate 5‐kinase is a putative target for increased intracellular phosphatidic acid

Determinants of specificity at the protein–lipid interface in membranes

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