The leucine 10 residue in the pleckstrin homology domain of ceramide kinase is crucial for its catalytic activity
- 19 July 2005
- journal article
- Published by Wiley in FEBS Letters
- Vol. 579 (20) , 4383-4388
- https://doi.org/10.1016/j.febslet.2005.06.079
Abstract
Ceramide kinase (CERK) converts ceramide (Cer) to ceramide-1-phosphate (C1P), a newly recognized bioactive molecule capable of regulating diverse cellular functions. The N-terminus of the CERK protein encompasses a sequence motif known as a pleckstrin homology (PH) domain. However, little is known regarding the functional roles of this domain in CERK. In this study, we have demonstrated that the PH domain of CERK is essential for its enzyme activity. Using site-directed mutagenesis, we have further determined that Leu10 in the PH domain has an important role in CERK activity. Replacing this residue with a neutral alanine or isoleucine, caused a dramatic decrease in CERK activity to 1% and 29%, respectively, compared to CERK, but had no effect on substrate affinity. The study presented here suggests that the PH domain of CERK is not only indispensable for its activity but also act as a regulator of CERK activityKeywords
This publication has 21 references indexed in Scilit:
- Asp177 in C4 domain of mouse sphingosine kinase 1a is important for the sphingosine recognitionFEBS Letters, 2004
- The role of sphingosine and ceramide kinases in inflammatory responsesImmunology Letters, 2004
- Conserved Amino Acid Residues in the COOH-terminal Tail Are Indispensable for the Correct Folding and Localization and Enzyme Activity of Neutral CeramidaseJournal of Biological Chemistry, 2004
- Ceramide Kinase Is a Mediator of Calcium-dependent Degranulation in Mast CellsJournal of Biological Chemistry, 2004
- Ceramide Kinase Mediates Cytokine- and Calcium Ionophore-induced Arachidonic Acid ReleaseJournal of Biological Chemistry, 2003
- Sphingosine-1-phosphate: an enigmatic signalling lipidNature Reviews Molecular Cell Biology, 2003
- N-terminal N -myristoylation of proteins: refinement of the sequence motif and its taxon-specific differences 1 1Edited by J. ThorntonJournal of Molecular Biology, 2002
- The Formation of Ceramide-1-phosphate during Neutrophil Phagocytosis and Its Role in Liposome FusionJournal of Biological Chemistry, 1998
- PH Domains: Diverse Sequences with a Common Fold Recruit Signaling Molecules to the Cell SurfaceCell, 1996
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970