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Hydrolytic and transphosphatidylation activities of phospholipase D from Savoy cabbage towards lysophosphatidylcholine

Virto, Carmen LU ; Svensson, Ingemar LU and Adlercreutz, Patrick LU (2000) In Chemistry and Physics of Lipids 106(1). p.41-51
Abstract

The hydrolysis and transphosphatidylation of lysophosphatidylcholine (LPC), with a partially purified preparation of phospholipase D (PL D) from Savoy cabbage, was investigated. These reactions were about 20 times slower than the hydrolysis of phosphatidylcholine (PC) in a micellar system. For the transfer reaction, 2 M glycerol was included in the media, which suppressed the hydrolytic reaction. Both reactions presented similar V(max) values, suggesting that the formation of the phosphatidyl-enzyme intermediate is the rate-limiting step. The enzyme had an absolute requirement for Ca2+, and the optimum concentration was approximately 40 mM CaCl2. K(Ca)(app) was calculated to be 8.6±0.74 mM for the hydrolytic and... (More)

The hydrolysis and transphosphatidylation of lysophosphatidylcholine (LPC), with a partially purified preparation of phospholipase D (PL D) from Savoy cabbage, was investigated. These reactions were about 20 times slower than the hydrolysis of phosphatidylcholine (PC) in a micellar system. For the transfer reaction, 2 M glycerol was included in the media, which suppressed the hydrolytic reaction. Both reactions presented similar V(max) values, suggesting that the formation of the phosphatidyl-enzyme intermediate is the rate-limiting step. The enzyme had an absolute requirement for Ca2+, and the optimum concentration was approximately 40 mM CaCl2. K(Ca)(app) was calculated to be 8.6±0.74 mM for the hydrolytic and 10±0.97 mM for the transphosphatidylation reaction. Both activities reached a maximum at pH 5.5, independent of Ca2+ concentration. Kinetic studies showed that the Km(app) for the glycerol in the transphosphatidylation reaction is 388±37 mM. Km(app) for the lysophosphatidylcholine depended on Ca2+ concentration and fell between 1 and 3 mM at CaCl2 concentrations from 4 to 40 mM. SDS, TX-100, and CTAB did not activate the enzyme as reported for phosphatidylcholine hydrolysis; on the contrary, reaction rates decreased at detergent concentrations at or above that of lysophosphatidylcholine. Copyright (C) 2000 Elsevier Science Ireland Ltd.

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type
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publication status
published
subject
keywords
Hydrolysis, Lysophosphatidic acid, Lysophosphatidylcholine, Lysophosphatidylglycerol, Phospholipase D, Transphosphatidylation
in
Chemistry and Physics of Lipids
volume
106
issue
1
pages
11 pages
publisher
Elsevier
external identifiers
  • scopus:0034213456
  • pmid:10878234
ISSN
0009-3084
DOI
10.1016/S0009-3084(00)00130-4
language
English
LU publication?
yes
id
f31915c6-ee6d-4368-a468-d75fe17a24a3
date added to LUP
2019-06-20 15:53:50
date last changed
2021-06-16 01:06:17
@article{f31915c6-ee6d-4368-a468-d75fe17a24a3,
  abstract     = {<p>The hydrolysis and transphosphatidylation of lysophosphatidylcholine (LPC), with a partially purified preparation of phospholipase D (PL D) from Savoy cabbage, was investigated. These reactions were about 20 times slower than the hydrolysis of phosphatidylcholine (PC) in a micellar system. For the transfer reaction, 2 M glycerol was included in the media, which suppressed the hydrolytic reaction. Both reactions presented similar V(max) values, suggesting that the formation of the phosphatidyl-enzyme intermediate is the rate-limiting step. The enzyme had an absolute requirement for Ca<sup>2+</sup>, and the optimum concentration was approximately 40 mM CaCl<sub>2</sub>. K(Ca)(app) was calculated to be 8.6±0.74 mM for the hydrolytic and 10±0.97 mM for the transphosphatidylation reaction. Both activities reached a maximum at pH 5.5, independent of Ca<sup>2+</sup> concentration. Kinetic studies showed that the Km(app) for the glycerol in the transphosphatidylation reaction is 388±37 mM. Km(app) for the lysophosphatidylcholine depended on Ca<sup>2+</sup> concentration and fell between 1 and 3 mM at CaCl<sub>2</sub> concentrations from 4 to 40 mM. SDS, TX-100, and CTAB did not activate the enzyme as reported for phosphatidylcholine hydrolysis; on the contrary, reaction rates decreased at detergent concentrations at or above that of lysophosphatidylcholine. Copyright (C) 2000 Elsevier Science Ireland Ltd.</p>},
  author       = {Virto, Carmen and Svensson, Ingemar and Adlercreutz, Patrick},
  issn         = {0009-3084},
  language     = {eng},
  month        = {06},
  number       = {1},
  pages        = {41--51},
  publisher    = {Elsevier},
  series       = {Chemistry and Physics of Lipids},
  title        = {Hydrolytic and transphosphatidylation activities of phospholipase D from Savoy cabbage towards lysophosphatidylcholine},
  url          = {http://dx.doi.org/10.1016/S0009-3084(00)00130-4},
  doi          = {10.1016/S0009-3084(00)00130-4},
  volume       = {106},
  year         = {2000},
}