Characterization of the kinetics of phospholipase C activity toward mixed micelles of sodium deoxycholate and dimyristoylphosphatidylcholine

Ranganathan, Radha; Tcacenco, Celize M.; Rosseto, Renato; Hajdu, Joseph

doi:10.1016/j.bpc.2006.02.012

Cited by 9 publications

(16 citation statements)

References 32 publications

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“…Thus, PLA 2 catalyzed cleavage results in increase of the fluorescence by the donor, and in loss of FRET (i.e., emission of the acceptor) that can be used to follow the hydrolysis by the enzyme. Along the same line, hydrolysis of the sn -2-ester bond in 27 causes an increase in fluorescence at 390 nm via de-quenching due to the departure of the paramagnetically labeled fatty acid quencher 22 from the substrate. Finally, we have applied the synthetic method to prepare compound 28 incorporating a chain-terminal ferrocene reporter group that was recently introduced as a redox-active label of phospholipids.…”

Section: Resultsmentioning

confidence: 92%

Synthesis of phospholipids on a glyceric acid scaffold: design and preparation of phospholipase A2 specific substrates

Rosseto

Hajdu

2014

Tetrahedron

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Synthesis of a new series of phospholipid analogues to serve as activity-based probes of secretory phospholipase A2 enzymes is reported. The synthesis is based upon 1) preparation of long-chain esters and amides of glyceric acid, followed by 2) regioselective derivatization of the diol function of the molecule to achieve phosphorylation at the primary hydroxyl group, and to introduce the incipient sn-2-ester group of the target compounds. The sequence has been shown to allow incorporation of fluorescent, paramagnetic, and redox-active reporter groups, leading to phospholipid analogues applicable to detect and measure enzyme activity, to develop highly specific, real-time spectroscopic assay of phospholipase A2 enzymes, as well as to track the metabolic fate of the hydrolysis products. The synthetic method has a great deal of flexibility to open the way to the design and synthesis of activity-probes for other phospholipid metabolizing enzymes as well.

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Section: Resultsmentioning

confidence: 92%

Synthesis of phospholipids on a glyceric acid scaffold: design and preparation of phospholipase A2 specific substrates

Rosseto

Hajdu

2014

Tetrahedron

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“…Bile salts are important biosurfactants, which exist in the living bodies of vertebrates [1][2][3] and often act as solubilizing or emulsifying agents for absorption of dietary lipids [4][5][6] or as gallstone solubilizing agents in clinical medicine. 7,8 All of them possess a rigid steroid backbone having polar hydroxyl groups on the concave R-face and methyl groups on the convex β-face.…”

Section: Introductionmentioning

confidence: 99%

Effect of Amino Acids on Aggregation Behaviors of Sodium Deoxycholate at Air/Water Surface: Surface Tension and Oscillating Bubble Studies

Pang

et al. 2010

Langmuir

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The aggregation behaviors of sodium deoxycholate (NaDC) at the air/water surface were investigated via surface tension and oscillating bubble measurements in the absence and presence of three alkaline amino acids, namely, L-Lysine (L-Lys), L-Arginine (L-Arg), and L-Histidine (L-His). The results of surface tension measurements show that NaDC has a lower ability to reduce the surface tension of water, because NaDC molecules orient at the surface in an oblique direction and tend to aggregate together, which is approved by molecular dynamics (MD) simulation. L-Lys is the most efficient of the three amino acids in reducing the critical aggregation concentration (cac) of NaDC in aqueous solution. The influence of amino acids on the dilational rheological properties of NaDC was studied using the drop shape analysis method in the frequency range from 0.02 to 0.5 Hz. The results reveal that the absolute modulus passes through a maximum value with increasing NaDC concentration. The addition of amino acids increases the absolute modulus of NaDC, and the maximum value is observed at much lower concentration. From the perspective of structures of amino acids, the performance of L-Arg is similar to that of L-His, and both of them bring out a smaller effect on the absolute modulus than that of L-Lys. From the above results, it may be presumed that electrostatic and hydrophobic effects are important impetus during the interaction between amino acids and NaDC at the air/water surface. Hydrogen bonding is so ubiquitous in the system that the difference of hydrogen bonding between NaDC and amino acid is ignored.

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“…On the other hand, cleavage of the sn-2-ester linkage of 2b leads to enhanced fluorescence at 397 nm via de-quenching, due to release of the paramagnetically labeled fatty acid from the molecule. 20 In conclusion, the synthesis here reported provides a facile and efficient method for the preparation of a new class of phospholipid analogues. The method should be applicable for the preparation of a wide range of functionalized phospholipid derivatives not only for development of new real-time spectrophotometric assays of phospholipases and highthroughput screening of phospholipase inhibitors, but also for the design and development of new membrane probes for the study of conformation and interaction of phospholipids in monolayers, bilayers and micelles.…”

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confidence: 79%

Synthesis of phosphatidylcholine analogues derived from glyceric acid: a new class of biologically active phospholipid compounds

Rosseto¹,

Tcacenco²,

Ranganathan³

et al. 2008

Tetrahedron Letters

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Synthesis of a new class of phosphatidylcholine analogues derived from glyceric acid is reported for spectroscopic studies of phospholipases and conformation of phospholipid side-chains in biological membranes, using fluorescence resonance energy transfer (FRET) techniques. Biologically active synthetic phospholipid compounds are required for structural and dynamic studies of biomembranes for establishment of structure-activity relationships with respect to phospholipid-phospholipid and phospholipid-protein interactions, as well as for mechanistic studies of phospholipid metabolizing enzymes. 1-3 Specifically, with the discovery that phospholipases generate a number of physiologically important lipid metabolites, including second messengers that are involved in cell signaling, 1 development of new synthetic methods for preparation of phospholipid derivatives became a key step in advancing membrane biochemistry. Phospholipid analogues incorporating spectroscopically active reporter groups have been shown to be valuable structural probes to study conformation and dynamics of phospholipids in aggregates (e.g. micelles, bilayers, and vesicles) as well as for development of spectroscopic assay systems of lipolytic enzymes, 4-6 and for in vivo monitoring the fate of products generated by phospholipids metabolizing enzymes. 7 As a part of our work in this area we focused on development of highly specific fluorescent phospholipase A 2 substrates for kinetic studies and for detection of in vivo activity of the enzyme. Phospholipases A 2 (PLA 2 , EC 3.1.1.4) comprise a large group of intracellular and secreted enzymes that catalyze the hydrolysis of the sn-2 ester function of glycerophospholipids to produce free fatty acids, such as arachidonic acid, and lysophospholipids. 8 Both products are precursors of signaling molecules with a multitude of biological functions. [8][9][10][11][12][13] Specifically,

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Characterization of the kinetics of phospholipase C activity toward mixed micelles of sodium deoxycholate and dimyristoylphosphatidylcholine

Cited by 9 publications

References 32 publications

Synthesis of phospholipids on a glyceric acid scaffold: design and preparation of phospholipase A2 specific substrates

Synthesis of phospholipids on a glyceric acid scaffold: design and preparation of phospholipase A2 specific substrates

Effect of Amino Acids on Aggregation Behaviors of Sodium Deoxycholate at Air/Water Surface: Surface Tension and Oscillating Bubble Studies

Synthesis of phosphatidylcholine analogues derived from glyceric acid: a new class of biologically active phospholipid compounds

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