Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D

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Standard

Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D. / Vinggaard, Anne Marie; Jensen, T.; Morgan, C.P.; Cockcroft, S.; Hansen, Harald S.

In: Biochemical Journal, Vol. 319, No. 3, 01.11.1996, p. 861-864.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Vinggaard, AM, Jensen, T, Morgan, CP, Cockcroft, S & Hansen, HS 1996, 'Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D', Biochemical Journal, vol. 319, no. 3, pp. 861-864.

APA

Vinggaard, A. M., Jensen, T., Morgan, C. P., Cockcroft, S., & Hansen, H. S. (1996). Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D. Biochemical Journal, 319(3), 861-864.

Vancouver

Vinggaard AM, Jensen T, Morgan CP, Cockcroft S, Hansen HS. Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D. Biochemical Journal. 1996 Nov 1;319(3):861-864.

Author

Vinggaard, Anne Marie ; Jensen, T. ; Morgan, C.P. ; Cockcroft, S. ; Hansen, Harald S. / Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D. In: Biochemical Journal. 1996 ; Vol. 319, No. 3. pp. 861-864.

Bibtex

@article{b50bf5e6f890415b92e81a6add068050,
title = "Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D",
abstract = "Phospholipase D (PLD) activity in crude or solubilized membranes from mammalian tissues is difficult to detect with the current assay techniques, unless a high radioactive concentration of substrate and/or long incubation times are employed. Generally, the enzyme has to be extracted and partially purified on one column before easy detection of activity. Furthermore, PLD activity in cultured cells can only be detected by the available assay techniques in the presence of guanosine 5'-[¿-thio]triphosphate (GTP[S]) and a cytosolic factor [usually ADP-ribosylation factor (Arf)]. In this paper we report that the use of didecanoyl phosphatidylcholine (C-PC) in mammalian PLD assays considerably increases the detection limit. C-PC was compared with the commonly used dipalmitoyl phosphatidylcholine (C-PC) as a substrate for PLD activity from membranes of human neutrophils, human placenta and pig brain, and from placental cytosol. C-PC was superior to C-PC by a factor of 2-28 depending on assay conditions and tissue, and it allowed the detection of GTP[S]- and Arf-stimulated PLD activity without addition of phosphatidylinositol 4,5-bisphosphate.",
author = "Vinggaard, {Anne Marie} and T. Jensen and C.P. Morgan and S. Cockcroft and Hansen, {Harald S.}",
year = "1996",
month = nov,
day = "1",
language = "English",
volume = "319",
pages = "861--864",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "3",

}

RIS

TY - JOUR

T1 - Didecanoyl phosphatidylcholine is a superior substrate for assaying mammalian phospholipase D

AU - Vinggaard, Anne Marie

AU - Jensen, T.

AU - Morgan, C.P.

AU - Cockcroft, S.

AU - Hansen, Harald S.

PY - 1996/11/1

Y1 - 1996/11/1

N2 - Phospholipase D (PLD) activity in crude or solubilized membranes from mammalian tissues is difficult to detect with the current assay techniques, unless a high radioactive concentration of substrate and/or long incubation times are employed. Generally, the enzyme has to be extracted and partially purified on one column before easy detection of activity. Furthermore, PLD activity in cultured cells can only be detected by the available assay techniques in the presence of guanosine 5'-[¿-thio]triphosphate (GTP[S]) and a cytosolic factor [usually ADP-ribosylation factor (Arf)]. In this paper we report that the use of didecanoyl phosphatidylcholine (C-PC) in mammalian PLD assays considerably increases the detection limit. C-PC was compared with the commonly used dipalmitoyl phosphatidylcholine (C-PC) as a substrate for PLD activity from membranes of human neutrophils, human placenta and pig brain, and from placental cytosol. C-PC was superior to C-PC by a factor of 2-28 depending on assay conditions and tissue, and it allowed the detection of GTP[S]- and Arf-stimulated PLD activity without addition of phosphatidylinositol 4,5-bisphosphate.

AB - Phospholipase D (PLD) activity in crude or solubilized membranes from mammalian tissues is difficult to detect with the current assay techniques, unless a high radioactive concentration of substrate and/or long incubation times are employed. Generally, the enzyme has to be extracted and partially purified on one column before easy detection of activity. Furthermore, PLD activity in cultured cells can only be detected by the available assay techniques in the presence of guanosine 5'-[¿-thio]triphosphate (GTP[S]) and a cytosolic factor [usually ADP-ribosylation factor (Arf)]. In this paper we report that the use of didecanoyl phosphatidylcholine (C-PC) in mammalian PLD assays considerably increases the detection limit. C-PC was compared with the commonly used dipalmitoyl phosphatidylcholine (C-PC) as a substrate for PLD activity from membranes of human neutrophils, human placenta and pig brain, and from placental cytosol. C-PC was superior to C-PC by a factor of 2-28 depending on assay conditions and tissue, and it allowed the detection of GTP[S]- and Arf-stimulated PLD activity without addition of phosphatidylinositol 4,5-bisphosphate.

UR - http://www.scopus.com/inward/record.url?scp=0029910470&partnerID=8YFLogxK

M3 - Journal article

AN - SCOPUS:0029910470

VL - 319

SP - 861

EP - 864

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 3

ER -

ID: 45563582