Phospholipase D Stimulates Release of Nascent Secretory Vesicles from the trans-Golgi Network
Ye-Guang Chen,Anirban Siddhanta,Cary D. Austin,Scott M. Hammond,Tsung Chang Sung,Michael A. Frohman,Andrew J. Morris,Dennis Shields +7 more
TLDR
It is demonstrated that immunoaffinity-purified human PLD1 stimulated nascent secretory vesicle budding from the TGN and ARF-1 stimulated endogenous PLD activity in Golgi membranes approximately threefold and this activation correlated with its enhancement of vesicles budding.Abstract:
Phospholipase D (PLD) is a phospholipid hydrolyzing enzyme whose activation has been implicated in mediating signal transduction pathways, cell growth, and membrane trafficking in mammalian cells. Several laboratories have demonstrated that small GTP-binding proteins including ADP-ribosylation factor (ARF) can stimulate PLD activity in vitro and an ARF-activated PLD activity has been found in Golgi membranes. Since ARF-1 has also been shown to enhance release of nascent secretory vesicles from the TGN of endocrine cells, we hypothesized that this reaction occurred via PLD activation. Using a permeabilized cell system derived from growth hormone and prolactin-secreting pituitary GH3 cells, we demonstrate that immunoaffinity-purified human PLD1 stimulated nascent secretory vesicle budding from the TGN approximately twofold. In contrast, a similarly purified but enzymatically inactive mutant form of PLD1, designated Lys898Arg, had no effect on vesicle budding when added to the permeabilized cells. The release of nascent secretory vesicles from the TGN was sensitive to 1% 1-butanol, a concentration that inhibited PLD-catalyzed formation of phosphatidic acid. Furthermore, ARF-1 stimulated endogenous PLD activity in Golgi membranes approximately threefold and this activation correlated with its enhancement of vesicle budding. Our results suggest that ARF regulation of PLD activity plays an important role in the release of nascent secretory vesicles from the TGN.read more
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A rac-like small G-protein from Brassica campestris activates a PKC-dependent phospholipase D.
TL;DR: Genomic analysis indicated that Brac1 belongs to a multigene family and is closely related to that of Arabidopsis thaliana Arac3 but shares relatively little homology with other members of the Ras superfamily.
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A low molecular weight factor from dividing cells activates phospholipase D in caveolin-enriched membrane microdomains.
Sergei Bychenok,David A. Foster +1 more
TL;DR: Evidence is provided for a highly conserved PLD-stimulating peptide that is elevated in response to mitogenic stimuli and was detected in a variety of rat tissues but was highest in testes, where a large percentage of cells are dividing.
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Phospholipase D2 Modulates the Secretory Pathway in RBL-2H3 Mast Cells
Claudia Maria Meirelles Marchini-Alves,Valeria Cintra Barbosa Lorenzi,Elaine Zayas Marcelino da Silva,Vivian Marino Mazucato,Maria Célia Jamur,Constance Oliver +5 more
TL;DR: Results suggest that PLD2 activity plays an important role in regulating glycoconjugate trafficking in mast cells and vesicular trafficking in secretory cells.
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Lipase activity in lipidomics - a hidden problem?
Samuel Furse,J.A. Killian +1 more
TL;DR: Protease inhibitors have been standard in proteomics research for some years and that for much of this time, convenient catch-all small-molecule inhibitor packages have been commercially available.
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