Nonspecific phospholipase C3 of radish has phospholipase D activity towards glycosylinositol phosphoceramide

Previously, we found an enzyme activity that produces phytoceramide 1-phosphate (PC1P) by hydrolysis of D position of glycosylinositol phosphoceramide (GIPC) in cabbage and called its activity as GIPC-phospholipase D (PLD). Here, we purified GIPC-PLD by sequential chromatography from radish roots. Peptide mass fingerprinting analysis revealed that the potential candidate for GIPC-PLD protein was nonspecific phospholipase C3 (NPC3), which has not been characterized as PLD. The recombinant NPC3 protein obtained by heterologous expression system with E. coli produced PC1P from GIPC, and showed essentially the same enzymatic properties as those we characterized as GIPC-PLD in cabbage, radish, and A. thaliana. From these results, we concluded that NPC3 is one of the enzymes that degrade GIPC, a major sphingolipid in plants.

This is the peer reviewed version of the following article: Hasi, R.Y., Ishikawa, T., Sunagawa, K., Takai, Y., Ali, H., Hayashi, J., Kawakami, R., Yuasa, K., Aihara, M., Kanemaru, K., Imai, H. and Tanaka, T. (2022), Nonspecific phospholipase C3 of radish has phospholipase D activity towards glycosylinositol phosphoceramide. FEBS Lett. 596: 3024-3036. which has been published in final form at https://doi.org/10.1002/1873-3468.14520. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.