Mammalian phospholipase D: activation by ammonium sulfate and nucleotides.

Nakamura, Shun‐ichi; Shimooku, Kiyoshi; Akisue, Toshihiro; Jinnai, Hitoshi; Hitomi, Tomohiro; Kiyohara, Yoshimoto; Ogino, Chiaki; Yoshida, Kohsuke; Nishizuka, Yasutomi

doi:10.1073/pnas.92.26.12319

Cited by 22 publications

(22 citation statements)

References 27 publications

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“…The formation of PtdEtOH was linear with the enzyme protein up to 50 ng per tube, and the reaction proceeded linearly with incubation time up to 60 min. This purification made PLD more sensitive to the cytosol factors for its activation than that previously reported [18].…”

Section: 3 Preparation Of Pldmentioning

confidence: 51%

“…The 36-kDa protein may differ from the 50-kDa protein in h ~matopoietic cell lines [13][14][15] 7 he 36-kDa protein containing fraction (1 mg protein) prepared as i~ the legend to Table 2 was applied to a Superose 12 column x~hich had been equilibrated with buffer A [18], and eluted with the s,tme buffer at a flow rate 0.5 ml/min using an FPLC system (Pharmacia). PtdEtOH produced (dpm x 10 -3) Supernatant and particulate fractions were prepared from various rat tissues [18].…”

Section: Discussionmentioning

confidence: 99%

“…An earlier report from this laboratory has described that under normal conditions membrane-bound PLD is in a latent form, but exhibits a high activity in the presence of ammonium sulfate and cytosol [18]. Using this enzyme assay, the kidney and spleen are shown to contain highest G-proteinsensitive PLD activity.…”

Section: Introductionmentioning

confidence: 99%

“…Proteins of the soluble fraction of rat kidney were precipitated with ammonium sulfate (30-70%), dissolved, dialyzed, and chromatographed on a Superdex 200 column as described earlier [18].…”

Section: Preparation Of Crude Cytosolmentioning

confidence: 99%

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Reconstitution of GTP‐γ‐S‐dependent phospholipase D activity with ARF, RhoA, and a soluble 36‐kDa protein

et al. 1996

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Section: 3 Preparation Of Pldmentioning

confidence: 51%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Preparation Of Crude Cytosolmentioning

confidence: 99%

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Reconstitution of GTP‐γ‐S‐dependent phospholipase D activity with ARF, RhoA, and a soluble 36‐kDa protein

et al. 1996

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“…The reaction was stopped by changing the medium to extraction buffer containing methanol, KCl, and EGTA. Phase separation, lipid extraction, and separation were carried out as described previously (14). PLD activity is expressed as a percentage of the radioactive […”

mentioning

confidence: 99%

Requirement of Phospholipase D for Ilimaquinone-induced Golgi Membrane Fragmentation

Sonoda

Okada

Jahangeer

et al. 2007

Journal of Biological Chemistry

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Although organelles such as the endoplasmic reticulum and Golgi apparatus are highly compartmentalized, these organelles are interconnected through a network of vesicular trafficking. The marine sponge metabolite ilimaquinone (IQ) is known to induce Golgi membrane fragmentation and is widely used to study the mechanism of vesicular trafficking. Although IQ treatment causes protein kinase D (PKD) activation, the detailed mechanism of IQ-induced Golgi membrane fragmentation remains unclear. In this work, we found that IQ treatment of cells caused a robust activation of phospholipase D (PLD). In the presence of 1-butanol but not 2-butanol, IQ-induced Golgi membrane fragmentation was completely blocked. In addition, IQ failed to induce Golgi membrane fragmentation in PLD knock-out DT40 cells. Furthermore, IQ-induced PKD activation was completely blocked by treatment with either 1-butanol or propranolol. Notably, IQ-induced Golgi membrane fragmentation was also blocked by propranolol treatment. These results indicate that PLD-catalyzed formation of phosphatidic acid is a prerequisite for IQ-induced Golgi membrane fragmentation and that enzymatic conversion of phosphatidic acid to diacylglycerol is necessary for subsequent activation of PKD and IQ-induced Golgi membrane fragmentation.Regulated vesicular trafficking is essential for the transport of proteins and lipids between subcellular compartments and for the maintenance of organelles in eukaryotic cells. These organelles help the cell establish an ordered structure within which its complex biochemical reactions and signaling processes are executed. The molecular mechanism underlying vesicular trafficking from the endoplasmic reticulum (ER) 2 to the cell surface through the Golgi apparatus has been investigated extensively. Malhotra and coworkers (1) have reported that the marine sponge metabolite ilimaquinone (IQ) inhibits protein trafficking events by reversibly breaking the Golgi apparatus into small vesicles. Several attempts were made to identify downstream molecules necessary for IQ-induced Golgi membrane vesiculation. Using a reconstitution assay system, protein kinase D (PKD) was identified as a key enzyme responsible for IQinduced Golgi membrane vesiculation (2). However, the exact mechanism underlying IQ-induced Golgi apparatus breakdown, especially a relationship between PKD and lipid remodeling, remains unclear. Membrane trafficking is energetically unfavorable and does not occur spontaneously in vivo, but occurs under strict control of specialized proteins. Evidence is accumulating that these proteins do not act alone but in concert with particular membrane lipids such as phosphoinositides (3), diacylglycerol (DAG) (4), and phosphatidic acid (PA) (5-7). Phospholipases hydrolyze phospholipids, the backbone of biological membranes. Phospholipase activity not only has a profound impact on the structure and stability of cellular membranes but also plays a pivotal role in regulating many critical cellular functions. Phospholipase D (PLD) generates PA, a mul...

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Inhibitory Effects of Thymus-Independent Type 2 Antigens on MHC Class II-Restricted Antigen Presentation: Comparative Analysis of Carbohydrate Structures and the Antigen Presenting Cell

González-Fernández

Carrasco-Marı́n

Álvarez-Domínguez

et al. 1997

Cellular Immunology

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Mammalian phospholipase D: activation by ammonium sulfate and nucleotides.

Abstract: ABSTRACT

Cited by 22 publications

References 27 publications

Reconstitution of GTP‐γ‐S‐dependent phospholipase D activity with ARF, RhoA, and a soluble 36‐kDa protein

Reconstitution of GTP‐γ‐S‐dependent phospholipase D activity with ARF, RhoA, and a soluble 36‐kDa protein

Requirement of Phospholipase D for Ilimaquinone-induced Golgi Membrane Fragmentation

Inhibitory Effects of Thymus-Independent Type 2 Antigens on MHC Class II-Restricted Antigen Presentation: Comparative Analysis of Carbohydrate Structures and the Antigen Presenting Cell

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