ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity

H. Alex Brown; Stephen Gutowski; Carolyn R. Moomaw; Clive Slaughter; Paul C. Sternwels

doi:10.1016/0092-8674(93)90323-I

ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity

H. Alex Brown, Stephen Gutowski, Carolyn R. Moomaw, Clive Slaughter, Paul C. Sternwels

Research output: Contribution to journal › Article › peer-review

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Abstract

The hydrolysis of phosphatidylcholine by phospholipase D (PLD) results in the production of phosphatidic acid and choline. An assay that uses an exogenous substrate was developed to measure this activity in membranes and solubilized preparations from HL60 cells. A cytosolic factor markedly enhanced PLD activity in membranes and was essential for GTPγS-dependent stimulation of an enriched preparation of PLD. The factor was purified to homogeneity from bovine brain cytosol and identified as a member of the ADP-Ribosylation Factor (ARF) subfamily of small G proteins. Subsequently, recombinant myristoylated ARF1 was found to be a better activator of PLD activity than was the nonmyristoylated form. ARF proteins have been implicated recently as factors for regulation of intracellular vesicle traffic. The current finding suggests that PLD activity plays a prominent role in the action of ARF and that ARF may be a key component in the generation of second messengers via phospholipase D.

Original language	English (US)
Pages (from-to)	1137-1144
Number of pages	8
Journal	Cell
Volume	75
Issue number	6
DOIs	https://doi.org/10.1016/0092-8674(93)90323-I
State	Published - Dec 17 1993

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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@article{ff8de82d94b747e8aa8c155eb528bc73,

title = "ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity",

abstract = "The hydrolysis of phosphatidylcholine by phospholipase D (PLD) results in the production of phosphatidic acid and choline. An assay that uses an exogenous substrate was developed to measure this activity in membranes and solubilized preparations from HL60 cells. A cytosolic factor markedly enhanced PLD activity in membranes and was essential for GTPγS-dependent stimulation of an enriched preparation of PLD. The factor was purified to homogeneity from bovine brain cytosol and identified as a member of the ADP-Ribosylation Factor (ARF) subfamily of small G proteins. Subsequently, recombinant myristoylated ARF1 was found to be a better activator of PLD activity than was the nonmyristoylated form. ARF proteins have been implicated recently as factors for regulation of intracellular vesicle traffic. The current finding suggests that PLD activity plays a prominent role in the action of ARF and that ARF may be a key component in the generation of second messengers via phospholipase D.",

author = "Brown, {H. Alex} and Stephen Gutowski and Moomaw, {Carolyn R.} and Clive Slaughter and Sternwels, {Paul C.}",

note = "Funding Information: We thank Dr. R. A. Kahn for the generous gift of recombinant ARFI (myr and non-myr) proteins and for his insightful comments on the manuscript. We also thank S. Afendis for excellent technical assistance in the sequencing of the ARF peptides, Dr. J. R. Hepler for many helpful conversations and comments on the manuscript, and K. Edwards for preparation of the manuscript. This investigation was supported by United States Public Health Service grant GM31954 and the Lucille P. Markey Charitable Trust.",

year = "1993",

month = dec,

day = "17",

doi = "10.1016/0092-8674(93)90323-I",

language = "English (US)",

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pages = "1137--1144",

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T1 - ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity

AU - Brown, H. Alex

AU - Gutowski, Stephen

AU - Moomaw, Carolyn R.

AU - Slaughter, Clive

AU - Sternwels, Paul C.

N1 - Funding Information: We thank Dr. R. A. Kahn for the generous gift of recombinant ARFI (myr and non-myr) proteins and for his insightful comments on the manuscript. We also thank S. Afendis for excellent technical assistance in the sequencing of the ARF peptides, Dr. J. R. Hepler for many helpful conversations and comments on the manuscript, and K. Edwards for preparation of the manuscript. This investigation was supported by United States Public Health Service grant GM31954 and the Lucille P. Markey Charitable Trust.

PY - 1993/12/17

Y1 - 1993/12/17

N2 - The hydrolysis of phosphatidylcholine by phospholipase D (PLD) results in the production of phosphatidic acid and choline. An assay that uses an exogenous substrate was developed to measure this activity in membranes and solubilized preparations from HL60 cells. A cytosolic factor markedly enhanced PLD activity in membranes and was essential for GTPγS-dependent stimulation of an enriched preparation of PLD. The factor was purified to homogeneity from bovine brain cytosol and identified as a member of the ADP-Ribosylation Factor (ARF) subfamily of small G proteins. Subsequently, recombinant myristoylated ARF1 was found to be a better activator of PLD activity than was the nonmyristoylated form. ARF proteins have been implicated recently as factors for regulation of intracellular vesicle traffic. The current finding suggests that PLD activity plays a prominent role in the action of ARF and that ARF may be a key component in the generation of second messengers via phospholipase D.

AB - The hydrolysis of phosphatidylcholine by phospholipase D (PLD) results in the production of phosphatidic acid and choline. An assay that uses an exogenous substrate was developed to measure this activity in membranes and solubilized preparations from HL60 cells. A cytosolic factor markedly enhanced PLD activity in membranes and was essential for GTPγS-dependent stimulation of an enriched preparation of PLD. The factor was purified to homogeneity from bovine brain cytosol and identified as a member of the ADP-Ribosylation Factor (ARF) subfamily of small G proteins. Subsequently, recombinant myristoylated ARF1 was found to be a better activator of PLD activity than was the nonmyristoylated form. ARF proteins have been implicated recently as factors for regulation of intracellular vesicle traffic. The current finding suggests that PLD activity plays a prominent role in the action of ARF and that ARF may be a key component in the generation of second messengers via phospholipase D.

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ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity

Abstract

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