Studies of Dlalkyl Ether Phospholipids. II. Requirement for a Liquid-Crystalline Substrate for Hydrolysis by Cabbage Leaf Phospholipase D

1971 ◽  
Vol 49 (12) ◽  
pp. 1362-1375 ◽  
Author(s):  
Jung-shou Chen ◽  
Peter G. Barton
Keyword(s):  
Diethyl Ether ◽  
Phospholipase D ◽  
Liquid Crystalline ◽  
Sodium Dodecyl ◽  
Crystalline Substrate ◽  
Hydrolytic Reaction ◽  
Hydrolysis Rates ◽  
Egg Lecithin ◽  
Ether Phospholipids ◽  
Hydrolysis Of

The hydrolysis of 1,2-ditetradecyl-, 1,2-dihexadecyl-, and 1,2-dioctadecyk-sn-glycero-3-phosphoryl-cholines by the soluble phospholipase D (EC 3.14.4) from cabbage leaf has been compared with that of egg lecithin. When a diethyl ether – chloroform mixture was used as activator low hydrolysis rates were observed for the dialkyl ether phospholipids because of their poor solubility in diethyl ether. Under these conditions the compounds did not inhibit the hydrolysis of egg lecithin. The hydrolytic reaction appeared to take place at the solvent–water interface resulting in a gradual denaturation of the enzyme. When sodium dodecyl sulfate was used as activator faster hydrolysis was evident due in part to a reduction in mesomorphic transition temperature (Tm) revealed by differential thermal analysis. Rates of hydrolysis of the homologous dialkyl ether phospholipids were inversely proportional to their Tm values in excess water. Under conditions required for effective hydrolysis of these compounds they inhibited the hydrolysis of egg lecithin. From studies of hydrolysis rates obtained with various combinations of phosphatidylcholine and phosphatidic acid analogues it was concluded that binding of enzyme to phospholipids and their subsequent hydrolysis required both an appropriate surface charge and a gel to liquid-crystalline phase transition in the substrate.

scholarly journals Crystal structure of thermostable alkylsulfatase SdsAP from Pseudomonas sp. S9

2017 ◽  
Vol 37 (3) ◽  
Author(s):  
Lifang Sun ◽  
Pu Chen ◽  
Yintao Su ◽  
Zhixiong Cai ◽  
Lingwei Ruan ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Sodium Dodecyl ◽  
Titration Calorimetry ◽  
Pseudomonas Sp ◽  
Sterol Carrier Protein ◽  
Dimerization Domain ◽  
Alkyl Chains ◽  
Hydrolysis Of

A novel alkylsulfatase from bacterium Pseudomonas sp. S9 (SdsAP) was identified as a thermostable alkylsulfatases (type III), which could hydrolyze the primary alkyl sulfate such as sodium dodecyl sulfate (SDS). Thus, it has a potential application of SDS biodegradation. The crystal structure of SdsAP has been solved to a resolution of 1.76 Å and reveals that SdsAP contains the characteristic metallo-β-lactamase-like fold domain, dimerization domain, and C-terminal sterol carrier protein type 2 (SCP-2)-like fold domain. Kinetic characterization of SdsAP to SDS by isothermal titration calorimetry (ITC) and enzymatic activity assays of constructed mutants demonstrate that Y246 and G263 are important residues for its preference for the hydrolysis of ‘primary alkyl’ chains, confirming that SdsAP is a primary alkylsulfatase.

scholarly journals The roles of phospholipase D and a GTP-binding protein in guanosine 5′-[γ-thio]triphosphate-stimulated hydrolysis of phosphatidylcholine in rat liver plasma membranes

1990 ◽  
Vol 272 (3) ◽  
pp. 749-753 ◽  
Author(s):  
K M Hurst ◽  
B P Hughes ◽  
G J Barritt
Keyword(s):  
Rat Liver ◽  
Phospholipase D ◽  
Plasma Membranes ◽  
Regulatory Protein ◽  
Gtp Binding ◽  
Liver Plasma Membranes ◽  
Low Concentrations ◽  
Rat Liver Plasma Membranes ◽  
Triton X ◽  
Hydrolysis Of

1. Guanosine 5′-[gamma-thio]triphosphate (GTP[S]) stimulated by 50% the rate of release of [3H]choline and [3H]phosphorylcholine in rat liver plasma membranes labelled with [3H]choline. About 70% of the radioactivity released in the presence of GTP[S] was [3H]choline and 30% was [3H]phosphorylcholine. 2. The hydrolysis of phosphorylcholine to choline and the conversion of choline to phosphorylcholine did not contribute to the formation of [3H]choline and [3H]phosphorylcholine respectively. 3. The release of [3H]choline from membranes was inhibited by low concentrations of SDS or Triton X-100. Considerably higher concentrations of the detergents were required to inhibit the release of [3H]phosphorylcholine. 4. Guanosine 5′-[beta gamma-imido]triphosphate and guanosine 5′-[alpha beta-methylene]triphosphate, but not adenosine 5′-[gamma-thio]-triphosphate, stimulated [3H]choline release to the same extent as did GTP[S]. The GTP[S]-stimulated [3H]choline release was inhibited by guanosine 5′-[beta-thio]diphosphate, GDP and GTP but not by GMP. 5. It is concluded that, in rat liver plasma membranes, (a) GTP[S]-stimulated hydrolysis of phosphatidylcholine is catalysed predominantly by phospholipase D with some contribution from phospholipase C, and (b) the stimulation of phosphatidylcholine hydrolysis by GTP[s] occurs via a GTP-binding regulatory protein.

scholarly journals Purification and Characterization of 2,6-β-d-Fructan 6-Levanbiohydrolase fromStreptomyces exfoliatus F3-2

2000 ◽  
Vol 66 (1) ◽  
pp. 252-256 ◽  
Author(s):  
Katsuichi Saito ◽  
Kazuya Kondo ◽  
Ichiro Kojima ◽  
Atsushi Yokota ◽  
Fusao Tomita
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Extracellular Enzyme ◽  
Molecular Weights ◽  
Sds Page ◽  
Monomeric Structure ◽  
Ph Range ◽  
Hydrolysis Of

ABSTRACT Streptomyces exfoliatus F3-2 produced an extracellular enzyme that converted levan, a β-2,6-linked fructan, into levanbiose. The enzyme was purified 50-fold from culture supernatant to give a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of this enzyme were 54,000 by SDS-PAGE and 60,000 by gel filtration, suggesting the monomeric structure of the enzyme. The isoelectric point of the enzyme was determined to be 4.7. The optimal pH and temperature of the enzyme for levan degradation were pH 5.5 and 60°C, respectively. The enzyme was stable in the pH range 3.5 to 8.0 and also up to 50°C. The enzyme gave levanbiose as a major degradation product from levan in an exo-acting manner. It was also found that this enzyme catalyzed hydrolysis of such fructooligosaccharides as 1-kestose, nystose, and 1-fructosylnystose by liberating fructose. Thus, this enzyme appeared to hydrolyze not only β-2,6-linkage of levan, but also β-2,1-linkage of fructooligosaccharides. From these data, the enzyme from S. exfoliatus F3-2 was identified as a novel 2,6-β-d-fructan 6-levanbiohydrolase (EC 3.2.1.64 ).

HYDROLYSIS OF LECITHIN BY PLANT PLASTID ENZYMES

1955 ◽  
Vol 33 (1) ◽  
pp. 575-589 ◽  
Author(s):  
Morris Kates
Keyword(s):  
Phosphatidic Acid ◽  
Inorganic Phosphate ◽  
Organic Phosphate ◽  
Water Soluble ◽  
Lag Phase ◽  
Carrot Root ◽  
First Order ◽  
Spinach Chloroplasts ◽  
Egg Lecithin ◽  
Hydrolysis Of

Enzymatic liberation of choline from egg lecithin by plastid fractions from sugar beet, spinach, and cabbage leaves and from carrot root was a rapid, first order reaction (up to 70% hydrolysis), and was not preceded by a lag phase. None of the choline-containing products of lecithin degradation (lysolecithin, glycerylphosphorylcholine, or phosphorylcholine) lost choline on incubation with spinach chloroplasts. Inorganic phosphate liberation from lecithin by the plastids was preceded by a lag phase and was much slower than choline liberation. Spinach chloroplasts catalyzed the liberation of inorganic phosphate from L-α-phosphatidic acid and from L-α-glycerophosphate. The water-soluble organic phosphate liberated from lecithin by spinach chloroplasts was identified chromatographically as phosphorylcholine. The ether-soluble organic phosphate produced during the hydrolysis of egg lecithin by carrot plastids was isolated and identified as L-α-phosphatidic acid. These observations suggest that the enzymatic hydrolysis of lecithin by plant plastids involves the following reactions: (1) lecithin → L-α-phosphatidic acid + choline; (2) L-α-phosphatidic acid → inorganic phosphate + diglyceride and/or (3) L-α-phosphatidic acid → glycerophosphate + fatty acids and (4) glycerophosphate → inorganic phosphate + glycerol; and (5) lecithin → phosphorylcholine + diglyceride. The L-α-structure for egg lecithin was confirmed.

Kinetics of alkaline hydrolysis of ethylp-nitrophenyl ethylphosphonate in the reverse micellar system: sodium dodecyl sulfate-hexanol-water

2000 ◽  
Vol 49 (2) ◽  
pp. 265-269
Author(s):  
L. Ya. Zakharova ◽  
F. G. Valeeva ◽  
L. A. Kudryavtseva ◽  
V. E. Bel'skii ◽  
E. P. Zhil'tsova ◽  
...  
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Alkaline Hydrolysis ◽  
Sodium Dodecyl ◽  
Micellar System ◽  
Dodecyl Sulfate ◽  
Reverse Micellar System ◽  
Kinetics Of ◽  
Hydrolysis Of ◽  
System Sodium ◽  
Reverse Micellar

scholarly journals Phorbol ester selectively stimulates the phospholipase D-mediated hydrolysis of phosphatidylethanolamine in multidrug-resistant MCF-7 human breast carcinoma cells

1994 ◽  
Vol 302 (3) ◽  
pp. 649-654 ◽  
Author(s):  
Z Kiss ◽  
M Tomono ◽  
W B Anderson
Keyword(s):  
Breast Carcinoma ◽  
Phospholipase D ◽  
Multidrug Resistant ◽  
Carcinoma Cells ◽  
Human Breast Carcinoma ◽  
Human Breast ◽  
Breast Carcinoma Cells ◽  
Human Breast Carcinoma Cells ◽  
Hydrolysis Of ◽  
Mcf 7

The phospholipase D (PLD)-mediated synthesis of phosphatidylethanol (PtdEtOH) and the hydrolysis of phosphatidylethanolamine (PtdEtn) and phosphatidylcholine (PtdCho) were examined in drug-sensitive and multidrug-resistant lines of MCF-7 human breast carcinoma cells. In drug-sensitive (MCF-7/WT) cells, the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) failed to enhance either the synthesis of PtdEtOH or the hydrolysis of either phospholipid. In the drug-resistant (MCF-7/MDR) cells, 100 nM PMA greatly enhanced both the synthesis of PtdEtOH (approximately 21-fold) and the hydrolysis of PtdEtn (approximately 29-fold), but had no effect on the hydrolysis of PtdCho. The PLD activators sphingosine and H2O2 were found to elicit only a slight (1.28-1.4-fold) stimulatory effect on PtdCho hydrolysis in both the MCF-7/WT and MCF-7/MDR cell types, and had only a small effect on PtdEtn hydrolysis in the MCF-7/WT cells as well. However, these agents significantly (approximately 2.6-3.5-fold) stimulated PtdEtn hydrolysis in the MCF-7/MDR cells. These data indicate that MCF-7/MDR cells contain a PtdEtn-specific PLD activity which can be selectively stimulated by PMA, sphingosine and H2O2.

Biosynthesis of glucosyl monophosphoryl undecaprenol and its role in lipoteichoic acid biosynthesis

1982 ◽  
Vol 152 (2) ◽  
pp. 616-625
Author(s):  
D J Mancuso ◽  
T H Chiu
Keyword(s):  
Thin Layer ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Lipoteichoic Acid ◽  
Molar Ratio ◽  
Streptococcus Sanguis ◽  
Chromatographic Profile ◽  
Hydrolysis Of ◽  
Cellulose Column

A glucophospholipid was detected in an incubation mixture containing UDP-glucose, MgCl2, ATP, and a particulate enzyme prepared from Streptococcus sanguis. The synthesis of this lipid was inhibited strongly by UDP and moderately by UMP. The molar ratio of glucose to phosphate in the purified lipid was found to be 1:1. Glucose and glucose 1-phosphate were released by mild alkaline hydrolysis of the glucophospholipid. The lipid produced by mild acid degradation of the purified lipid yielded a thin-layer chromatographic profile similar to that of acid-treated undecaprenol. One of the minor components exhibited the same mobility as untreated undecaprenol. To characterize further the lipid moiety of the glucophospholipid, a polyisoprenol was purified from the neutral lipid of S. sanguis. The polyisoprenol was converted in the presence of ATP, UDP-glucose, and the particulate enzyme into a lipid which exhibited the same thin-layer chromatographic mobility as the glucophospholipid. The structure of the polyisoprenol was determined by nuclear magnetic resonance and mass spectrometry to be an undecaprenol with an internal cis-trans ratio of 7:2. These results indicate that the glucophospholipid is glucosyl monophosphoryl undecaprenol. The glucosyl moiety of the glucophospholipid was shown to be incorporated in the presence of the particulate enzyme into a macromolecule which was characterized as a lipoteichoic acid by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and DEAE-cellulose column chromatography. This result indicates that glucosyl monophosphoryl undecaprenol is the direct glucosyl donor in the synthesis of lipoteichoic acid.

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