Evidence for and Characterization of Ca2+ Binding to the Catalytic Region of Arabidopsis thaliana Phospholipase Dβ

Pappan, Kirk L.; Zheng, Li; Krishnamoorthi, Ramaswamy; Wang, Xuemin

doi:10.1074/jbc.m402789200

Cited by 31 publications

(24 citation statements)

References 41 publications

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“…The bacteria were grown to an OD 600 of 0.7 and induced with 0.1 mM isopropyl 1-thio-b-D-galactopyranoside for 16 h at 168C. The pPLAIIIb-His fusion protein was purified as described previously (Pappan et al, 2004). Briefly, the bacterial pellet was resuspended in STE buffer containing 1 mg/mL lysozyme (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 1 mM EDTA).…”

Section: Methods Pplaiiib Cloning and Protein Purification From Eschementioning

confidence: 99%

Patatin-Related Phospholipase pPLAIIIβ-Induced Changes in Lipid Metabolism Alter Cellulose Content and Cell Elongation in Arabidopsis

et al. 2011

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The release of fatty acids from membrane lipids has been implicated in various plant processes, and the patatin-related phospholipases (pPLAs) constitute a major enzyme family that catalyzes fatty acid release. The Arabidopsis thaliana pPLA family has 10 members that are classified into three groups. Group 3 pPLAIII has four members but lacks the canonical lipase/esterase consensus catalytic sequences, and their enzymatic activity and cellular functions have not been delineated. Here, we show that pPLAIIIb hydrolyzes phospholipids and galactolipids and additionally has acyl-CoA thioesterase activity. Alterations of pPLAIIIb result in changes in lipid levels and composition. pPLAIIIb-KO plants have longer leaves, petioles, hypocotyls, primary roots, and root hairs than wild-type plants, whereas pPLAIIIb-OE plants exhibit the opposite phenotype. In addition, pPLAIIIb-OE plants have significantly lower cellulose content and mechanical strength than wild-type plants. Root growth of pPLAIIIb-KO plants is less sensitive to treatment with free fatty acids, the enzymatic products of pPLAIIIb, than wild-type plants; root growth of pPLAIIIb-OE plants is more sensitive. These data suggest that alteration of pPLAIIIb expression and the resulting lipid changes alter cellulose content and cell elongation in Arabidopsis.

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Section: Methods Pplaiiib Cloning and Protein Purification From Eschementioning

confidence: 99%

Patatin-Related Phospholipase pPLAIIIβ-Induced Changes in Lipid Metabolism Alter Cellulose Content and Cell Elongation in Arabidopsis

et al. 2011

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“…The bacteria were grown to an optical density at 600 nm of 0.7 and induced with 0.1 mM isopropyl 1-thio-b-D-galactopyranoside for 16 h at 16°C. The pPLAIIId-63His fusion protein was purified as described previously (Pappan et al, 2004). Briefly, the bacterial pellet was resuspended in STE buffer containing 1 mg mL 21 lysozyme (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 1 mM EDTA).…”

Section: Analysis Of Fatty Acid Composition and Oil Contentmentioning

confidence: 99%

Patatin-Related Phospholipase pPLAIIIδ Increases Seed Oil Content with Long-Chain Fatty Acids in Arabidopsis

Bahn

Fan

et al. 2013

Plant Physiology

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The release of fatty acids from membrane lipids has been implicated in various metabolic and physiological processes, but in many cases, the enzymes involved and their functions in plants remain unclear. Patatin-related phospholipase As (pPLAs) constitute a major family of acyl-hydrolyzing enzymes in plants. Here, we show that pPLAIIId promotes the production of triacylglycerols with 20-and 22-carbon fatty acids in Arabidopsis (Arabidopsis thaliana). Of the four pPLAIIIs (a, b, g, d), only pPLAIIId gene knockout results in a decrease in seed oil content, and pPLAIIId is most highly expressed in developing embryos. The overexpression of pPLAIIId increases the content of triacylglycerol and 20-and 22-carbon fatty acids in seeds with a corresponding decrease in 18-carbon fatty acids. Several genes in the glycerolipid biosynthetic pathways are up-regulated in pPLAIIId-overexpressing siliques. pPLAIIId hydrolyzes phosphatidylcholine and also acyl-coenzyme A to release fatty acids. pPLAIIId-overexpressing plants have a lower level, whereas pPLAIIId knockout plants have a higher level, of acyl-coenzyme A than the wild type. Whereas seed yield decreases in transgenic plants that ubiquitously overexpress pPLAIIId, seed-specific overexpression of pPLAIIId increases seed oil content without any detrimental effect on overall seed yield. These results indicate that pPLAIIId-mediated phospholipid turnover plays a role in fatty acid remodeling and glycerolipid production.Lipids play essential structural, metabolic, and regulatory roles in plant growth, development, and stress responses. In addition, plant lipids are a major source of food and renewable materials for various industrial and energy applications (Dyer et al., 2008;Hayden

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“…The above-described amino acid substitutions are responsible for the low affinity of PLDα and PLDε for Ca 2+ , and therefore these isoenzymes hydrolyze phospholipids only in the presence of high concentrations of Ca 2+ [2] or at low pH values increasing the affinity for calcium. The Ca 2+ binding has been also observed in the active site of PLDα [60] and PLDβ, where it is stimulated by phosphatidylserine [61].…”

Section: Specific Features Of Primary Structure Of Pldmentioning

confidence: 86%

“…This leads to conformational changes in the enzymes that result in strengthening the substrate binding and weakening the association with PtdIns(4,5)P 2 [2]. Calcium also binds with the active site of PLDβ, which increases its affinity for the activator (PtdIns(4,5)P 2 ) as discriminated for the affinity for the substrate (phosphatidylcholine) [61]. In turn, the binding of PtdIns(4,5)P 2 with the active site of the enzyme increases its affinity for the substrate [2].…”

Section: Regulatory Mechanisms Of Pld Activitymentioning

confidence: 99%

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Molecular structure of phospholipase D and regulatory mechanisms of its activity in plant and animal cells

Kolesnikov

Nokhrina

Kretynin

et al. 2012

Biochemistry Moscow

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Abbreviations: Gα, α-subunit of heterotrimeric G-protein; PKC, protein kinase C; PLD, phospholipase D; PtdIns(4,5)P 2 , phosphatidylinositol-4,5-bisphosphate. * To whom correspondence should be addressed.Abstract⎯Phospholipase D (PLD) catalyzes hydrolysis of phospholipids with production of phosphatidic acids, which often act as secondary messengers on transmission of intracellular signals. This review summarizes data of various leading laboratories on specific features of organization and regulation of PLD activity in plant and animal cells. The main structural domains of PLD (C2, PX, PH), the active site, and other functionally important parts of the enzyme are considered. Regulatory mechanisms of PLD activity are characterized in detail. Studies associated with molecular design, analysis, and synthesis of new nontoxic substances capable of inhibiting different PLD isoenzymes in vivo are shown to be promising for biotechnology and medicine.

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Evidence for and Characterization of Ca2+ Binding to the Catalytic Region of Arabidopsis thaliana Phospholipase Dβ

Cited by 31 publications

References 41 publications

Patatin-Related Phospholipase pPLAIIIβ-Induced Changes in Lipid Metabolism Alter Cellulose Content and Cell Elongation in Arabidopsis

Patatin-Related Phospholipase pPLAIIIβ-Induced Changes in Lipid Metabolism Alter Cellulose Content and Cell Elongation in Arabidopsis

Patatin-Related Phospholipase pPLAIIIδ Increases Seed Oil Content with Long-Chain Fatty Acids in Arabidopsis

Molecular structure of phospholipase D and regulatory mechanisms of its activity in plant and animal cells

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