Phospholipase Dalpha from sunflower (Helianthus annuus): cloning and functional characterization

Journal of Plant Physiology
A J Moreno-PérezJ J Salas

Abstract

D type phospholipases (PLD) are enzymes that hydrolyze the head group of phospholipids to produce phosphatidic acid. This activity is ubiquitous in plant tissues, and has been isolated and characterized from different species and organs. Several families of these proteins have been described in plants on the basis of their gene sequences (PLD alpha, beta, gamma, delta, zeta and epsilon). They have been shown to be involved in many metabolic events, such as response to abiotic stress, signal transduction, and membrane lipid turnover and degradation. In the present study, PLD activity was measured in the soluble fractions isolated from different organs of this plant. A PLD of alpha type was cloned from leaf cDNA that was responsible for most of this activity. The gene encoding this 810 aa protein was heterologously expressed in E. coli. This protein was not lethal for the eukaryotic host, although it altered its phospholipid profile. PLDalpha was purified to almost homogeneity by His-tag affinity chromatography, displaying an optimum pH of 6.5 and strong dependence on the presence of Ca(2+) and SDS in the assay medium. The enzyme was active towards phosphatidylcholine, Phosphatidylethanolamine and phosphatidylglycerol. Furthermor...Continue Reading

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Citations

Feb 22, 2012·Biochemistry. Biokhimii︠a︡·Y S KolesnikovV S Kravets
Nov 3, 2011·Lipids in Health and Disease·Slim Abdelkafi, Abdelkarim Abousalham
Jun 10, 2015·Phytochemistry·Hafeeza KhatoonRenate Ulbrich-Hofmann
Jan 20, 2017·BioMed Research International·Renaud RahierAbdelkarim Abousalham
May 15, 2021·Critical Reviews in Biotechnology·Peng ZhangJin-Song Shi

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