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Bioorganic & Medicinal Chemistry May 2023In this work, we report the design, synthesis, and application of a bis-pyrene phospholipid probe for detection of phospholipase A action through changes in pyrene...
In this work, we report the design, synthesis, and application of a bis-pyrene phospholipid probe for detection of phospholipase A action through changes in pyrene monomer and excimer fluorescence intensities. Continuous fluorometric assays enabled detection of the activities of multiple PLA enzymes as well as the decrease in catalysis by PLA from honey bee venom caused by the inhibitor p-bromo phenacylbromide. Thin-layer chromatography and mass spectrometry analysis were also used to validate probe hydrolysis by PLA. Mass spectrometry data also supported cleavage of the probe by phospholipase C and D enzymes, although changes in fluorescence were not observed in these cases. Nevertheless, the bis-pyrene phospholipid probe developed in this work is effective for detection of PLA enzyme activity through an assay that enables screening for inhibitor development.
Topics: Hydrolysis; Phospholipases; Phospholipases A2; Phospholipids; Pyrenes
PubMed: 37150117
DOI: 10.1016/j.bmc.2023.117301 -
Annals of the Rheumatic Diseases Nov 1989
Topics: Arthritis, Rheumatoid; Humans; Inflammation; Phospholipases; Phospholipases A; Phospholipases A2; Synovial Fluid
PubMed: 2596887
DOI: 10.1136/ard.48.11.962-b -
Seikagaku. the Journal of Japanese... Dec 1987
Review
Topics: Animals; Blood Platelets; Phospholipases; Phospholipases A; Phospholipases A2
PubMed: 3329671
DOI: No ID Found -
Biochemistry Apr 2021Phospholipase A/acyltransferase 3 (PLAAT3) and PLAAT4 are enzymes involved in the synthesis of bioactive lipids. Despite sequential and structural similarities, the two...
Phospholipase A/acyltransferase 3 (PLAAT3) and PLAAT4 are enzymes involved in the synthesis of bioactive lipids. Despite sequential and structural similarities, the two enzymes differ in activity and specificity. The relation between the activity and dynamics of the N-terminal domains of PLAAT3 and PLAAT4 was studied. PLAAT3 has a much higher melting temperature and exhibits less nanosecond and millisecond dynamics in the active site, in particular in loop L2(B6), as shown by NMR spectroscopy and molecular dynamics calculations. Swapping the L2(B6) loops between the two PLAAT enzymes results in strongly increased phospholipase activity in PLAAT3 but no reduction in PLAAT4 activity, indicating that this loop contributes to the low activity of PLAAT3. The results show that, despite structural similarity, protein dynamics differ substantially between the PLAAT variants, which can help to explain the activity and specificity differences.
Topics: Catalytic Domain; Molecular Dynamics Simulation; Phospholipases; Substrate Specificity; Temperature
PubMed: 33749246
DOI: 10.1021/acs.biochem.0c00974 -
Analytical Biochemistry Dec 2006
Topics: Attention; Enzyme Inhibitors; Hydrolysis; Inflammation; Inhibition, Psychological; Liposomes; Phospholipases; Phospholipases A; Phospholipases A2; Phospholipids; Substrate Specificity
PubMed: 17045950
DOI: 10.1016/j.ab.2006.08.031 -
Analytical Biochemistry Jan 1963
Topics: Chemistry Techniques, Analytical; Egg Yolk; Nephelometry and Turbidimetry; Phospholipases; Type C Phospholipases
PubMed: 13940740
DOI: 10.1016/0003-2697(63)90055-1 -
Ceskoslovenska Fysiologie Sep 1998Phospholipase A2 (PLA2) hydrolyses membrane phospholipids (PL) and it may release arachidonic acid (AA)--the precursor of eicosanoids--from the sn-2 position. PLA2 and... (Review)
Review
Phospholipase A2 (PLA2) hydrolyses membrane phospholipids (PL) and it may release arachidonic acid (AA)--the precursor of eicosanoids--from the sn-2 position. PLA2 and metabolites of its catalytic activity participate in many processes in the organism: metabolism of lipids, inflammation and immune reactions, membrane and tissue reparation, proliferation, and others. PLA2 as also an important element in the signal transduction. In the present article, PLA2 is characterised, classified into several types, and its mechanism of action together with its possible role in the disease processes are described. The attention is aimed at two forms of PLA2: The secretory (PLA2) of the type II which is associated with the inflammation injury, and the cytosolic PLA2 which is the main catalyst in the liberation of AA and which participates in the signal transduction. Other forms of PLA2 has been also described.
Topics: Arachidonic Acid; Inflammation; Phospholipases A; Phospholipases A2; Signal Transduction
PubMed: 9748759
DOI: No ID Found -
Advances in Immunology 2001
Review
Topics: Animals; Humans; Mammals; Phospholipases A; Receptors, Cell Surface; Receptors, Phospholipase A2
PubMed: 11293116
DOI: 10.1016/s0065-2776(01)77017-4 -
Tanpakushitsu Kakusan Koso. Protein,... Nov 1976
Review
Topics: Animals; Bacterial Toxins; Capillary Permeability; Chromatography, Affinity; Chromatography, Ion Exchange; Clostridium perfringens; Culture Media; Guinea Pigs; Hemolysis; Microcirculation; Molecular Weight; Phospholipases; Rats
PubMed: 194279
DOI: No ID Found -
ACS Chemical Biology Mar 2006Quantitative real-time in situ activity assays are necessary for determining the physiological function and regulation of enzymes. A paper in this issue reports the... (Review)
Review
Quantitative real-time in situ activity assays are necessary for determining the physiological function and regulation of enzymes. A paper in this issue reports the synthesis of a series of new fluorogenic phospholipids that allow fast real-time measurements of cellular activity and head group selectivity of an important family of enzyme, phospholipases.
Topics: Computer Systems; Enzyme Activation; Intracellular Fluid; Phospholipases A; Phospholipases A2
PubMed: 17163643
DOI: 10.1021/cb600079t