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A Novel Cytosolic Calcium-independent Phospholipase A2 Contains Eight Ankyrin Motifs
Open accessAbstract
We report the purification, molecular cloning, and expression of a novel cytosolic calcium-independent phospholipase A2 (iPLA2) from Chinese hamster ovary cells, which lacks extended homology to other phospholipases. iPLA2 is an 85-kDa protein that exists as a multimeric complex of 270-350 kDa with a specific activity of 1 micromol/min/mg. The full-length cDNA clone encodes a 752-amino acid cytoplasmic protein with one lipase motif (GXS465XG) and eight ankyrin repeats. Expression of the cDNA in mammalian cells generates an active 85-kDa protein. Mutagenesis studies show that Ser465 and the ankyrin repeats are required for activity. We demonstrate that iPLA2 selectively hydrolyzes the sn-2 over sn-1 fatty acid by 5-fold for 1,2-dipalmitoyl phosphatidylcholine in a mixed micelle. Moreover, we found the fatty acid preference at the sn-2 position to be highly dependent upon substrate presentation. However, iPLA2 does have a marked preference for 1,2-dipalmitoyl phosphatidic acid presented in a vesicle, generating the lipid second messenger lysophosphatidic acid. Finally the enzyme is able to hydrolyze the acetyl moiety at the sn-2 position of platelet-activating factor.
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Clark, J. D., Lin, L.-L., Kriz, R. W., Ramesha, C. S., Sultzman, L. A., Lin, A. Y., Milona, N., & Knopf, J. L. (1991) · Cell · doi:10.1016/0092-8674(91)90556-e · Cited by 1,688
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Cited by
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Lipolysis – A highly regulated multi-enzyme complex mediates the catabolism of cellular fat stores
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Balsinde, J., Winstead, M. V., & Dennis, E. A. (2002) · FEBS Letters · doi:10.1016/s0014-5793(02)03413-0 · Cited by 477
Chen, J.-W., Dodia, C., Feinstein, S. I., Jain, M. K., & Fisher, A. B. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m005073200 · Cited by 447
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Metadata
- Authors
- Tang, J., Kriz, R. W., Wolfman, N., Shaffer, M., Seehra, J., & Jones, S. S.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1997-03
- Volume
- 272
- Issue
- 13
- Pages
- 8567-8575
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2069404833
- Cited by
- 299
- References
- 1
- FWCI
- 14.23
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Tang, J., Kriz, R. W., Wolfman, N., Shaffer, M., Seehra, J., & Jones, S. S. (1997). A Novel Cytosolic Calcium-independent Phospholipase A2 Contains Eight Ankyrin Motifs. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.272.13.8567
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