Noiriel_2004_Eur.J.Biochem_271_3752

Reference

Title : Expression in yeast of a novel phospholipase A1 cDNA from Arabidopsis thaliana - Noiriel_2004_Eur.J.Biochem_271_3752
Author(s) : Noiriel A , Benveniste P , Banas A , Stymne S , Bouvier-Nave P
Ref : European Journal of Biochemistry , 271 :3752 , 2004
Abstract :

During a search for cDNAs encoding plant sterol acyltransferases, we isolated four full-length cDNAs from Arabidopsis thaliana that encode proteins with substantial identity with animal lecithin : cholesterol acyltransferases (LCATs). The expression of one of these cDNAs, AtLCAT3 (At3g03310), in various yeast strains resulted in the doubling of the triacylglycerol content. Furthermore, a complete lipid analysis of the transformed wild-type yeast showed that its phospholipid content was lower than that of the control (void plasmid-transformed) yeast whereas lysophospholipids and free fatty acids increased. When microsomes from the AtLCAT3-transformed yeast were incubated with di-[1-14C]oleyl phosphatidylcholine, both the lysophospholipid and free fatty acid fractions were highly and similarly labelled, whereas the same incubation with microsomes from the control yeast produced a negligible labelling of these fractions. Moreover when microsomes from AtLCAT3-transformed yeast were incubated with either sn-1- or sn-2-[1-14C]acyl phosphatidylcholine, the distribution of the labelling between the free fatty acid and the lysophosphatidylcholine fractions strongly suggested a phospholipase A1 activity for AtLCAT3. The sn-1 specificity of this phospholipase was confirmed by gas chromatography analysis of the hydrolysis of 1-myristoyl, 2-oleyl phosphatidylcholine. Phosphatidylethanolamine and phosphatidic acid were shown to be also hydrolysed by AtLCAT3, although less efficiently than phosphatidylcholine. Lysophospatidylcholine was a weak substrate whereas tripalmitoylglycerol and cholesteryl oleate were not hydrolysed at all. This novel A. thaliana phospholipase A1 shows optimal activity at pH 6-6.5 and 60-65 degrees C and appears to be unaffected by Ca2+. Its sequence is unrelated to all other known phospholipases. Further studies are in progress to elucidate its physiological role.

PubMedSearch : Noiriel_2004_Eur.J.Biochem_271_3752
PubMedID: 15355352
Gene_locus related to this paper: arath-At5g13640 , arath-LCAT1 , arath-LCAT4 , arath-LCAT3 , lyces-q71lx7 , medtr-q6xsh3 , medtr-Q8GZM5 , medtr-q71ce5 , tobac-q71lw1

Related information

Gene_locus arath-At5g13640    arath-LCAT1    arath-LCAT4    arath-LCAT3    lyces-q71lx7    medtr-q6xsh3    medtr-Q8GZM5    medtr-q71ce5    tobac-q71lw1

Citations formats

Noiriel A, Benveniste P, Banas A, Stymne S, Bouvier-Nave P (2004)
Expression in yeast of a novel phospholipase A1 cDNA from Arabidopsis thaliana
European Journal of Biochemistry 271 :3752

Noiriel A, Benveniste P, Banas A, Stymne S, Bouvier-Nave P (2004)
European Journal of Biochemistry 271 :3752