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.
(Less)
- author
- Virto, Carmen
LU
; Svensson, Ingemar
LU
and Adlercreutz, Patrick
LU
- organization
- publishing date
- 2000-06-01
- type
- Contribution to journal
- 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},
}