What question did this study set out to answer?

To purify phosphatidylcholine-specific phospholipase D from pig lung microsomes to homogeneity and determine its catalytic and regulatory properties.

December 1, 1994Journal of Biological ChemistryOpen Access

Purification and characterization of phosphatidylcholine phospholipase D from pig lung.

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Authors

SOS. OkamuraUniversity of Ferrara SYSatoshi YamashitaTaiho Pharmaceutical (Japan)

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Implication

Biochemical study demonstrates the purification of a 190-kDa phospholipase D in pig lung microsomes, revealing strict phosphatidylcholine specificity and fatty acid regulation.

Key Points

To purify phosphatidylcholine-specific phospholipase D from pig lung microsomes to homogeneity and determine its catalytic and regulatory properties.
Solubilized microsomal phospholipase D from pig lung using heptylthioglucoside detergent.
Purified the enzyme sequentially across six chromatographic steps: sulfate-Cellulofine, ether-Toyopearl, chelate-Toyopearl, Q-Sepharose, heparin-Toyopearl, and hydroxyapatite.
Assayed substrate specificity, optimum pH, kinetic parameters, cation dependence, and modulation by fatty acids and detergents.
Purified the enzyme 2,200-fold to homogeneity, yielding a single protein band with a molecular weight of 190,000 on SDS-PAGE.
Identified an optimum pH of 6.6 and half-maximal activity at 0.8 mM dipalmitoylglycerophosphocholine, with absolute substrate selectivity for phosphatidylcholine over lysophosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol.
Demonstrated generation of phosphatidic acid and choline, production of phosphatidylethanol in the presence of ethanol, and strong enzyme stimulation by unsaturated fatty acids combined with magnesium ions.

Cite This Study

Okamura et al. (1994) studied this question.

synapsesocial.com/papers/6a85598b734852141cd4c929 https://doi.org/10.1016/s0021-9258(18)47410-8

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Implication

Biochemical study demonstrates the purification of a 190-kDa phospholipase D in pig lung microsomes, revealing strict phosphatidylcholine specificity and fatty acid regulation.

Purification and characterization of phosphatidylcholine phospholipase D from pig lung.

Discussion

Member takes

Implication

Key Points

Cite This Study

Purification and characterization of phosphatidylcholine phospholipase D from pig lung.

Discussion

Member takes

Implication

Key Points

Cite This Study

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