Functional Characterization of the N-Terminal C2 Domain from Arabidopsis thaliana Phospholipase Dα and Dβ
Joint Authors
Rahier, Renaud
Noiriel, Alexandre
Abousalham, Abdelkarim
Source
Issue
Vol. 2016, Issue 2016 (31 Dec. 2016), pp.1-15, 15 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-12-22
Country of Publication
Egypt
No. of Pages
15
Main Subjects
Abstract EN
Most of plant phospholipases D (PLD) exhibit a C2-lipid binding domain of around 130 amino acid residues at their N-terminal region, involved in their Ca2+-dependent membrane binding.
In this study, we expressed and partially purified catalytically active PLDα from Arabidopsis thaliana (AtPLDα) in the yeast Pichia pastoris.
The N-terminal amino acid sequence of the recombinant AtPLDα was found to be NVEETIGV and thus to lack the first 35 amino acid belonging to the C2 domain, as found in other recombinant or plant purified PLDs.
To investigate the impact of such a cleavage on the functionality of C2 domains, we expressed, in E.
coli, purified, and refolded the mature-like form of the C2 domain of the AtPLDα along with its equivalent C2 domain of the AtPLDβ, for the sake of comparison.
Using Förster Resonance Energy Transfer and dot-blot assays, both C2 domains were shown to bind phosphatidylglycerol in a Ca2+-independent manner while phosphatidic acid and phosphatidylserine binding were found to be enhanced in the presence of Ca2+.
Amino acid sequence alignment and molecular modeling of both C2 domains with known C2 domain structures revealed the presence of a novel Ca2+-binding site within the C2 domain of AtPLDα.
American Psychological Association (APA)
Rahier, Renaud& Noiriel, Alexandre& Abousalham, Abdelkarim. 2016. Functional Characterization of the N-Terminal C2 Domain from Arabidopsis thaliana Phospholipase Dα and Dβ. BioMed Research International،Vol. 2016, no. 2016, pp.1-15.
https://search.emarefa.net/detail/BIM-1097143
Modern Language Association (MLA)
Rahier, Renaud…[et al.]. Functional Characterization of the N-Terminal C2 Domain from Arabidopsis thaliana Phospholipase Dα and Dβ. BioMed Research International No. 2016 (2016), pp.1-15.
https://search.emarefa.net/detail/BIM-1097143
American Medical Association (AMA)
Rahier, Renaud& Noiriel, Alexandre& Abousalham, Abdelkarim. Functional Characterization of the N-Terminal C2 Domain from Arabidopsis thaliana Phospholipase Dα and Dβ. BioMed Research International. 2016. Vol. 2016, no. 2016, pp.1-15.
https://search.emarefa.net/detail/BIM-1097143
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1097143