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Post-translational modification regulates prostaglandin D2 synthase apoptotic activity: Characterization by site-directed mutagenesis
- Source :
- Prostaglandins & Other Lipid Mediators. 83:25-32
- Publication Year :
- 2007
- Publisher :
- Elsevier BV, 2007.
-
Abstract
- Lipocalin-type prostaglandin D(2) synthase (L-PGDS) is a highly glycosylated protein found in several body fluids. Elevated L-PGDS levels have been observed in the serum of patients with renal impairment, diabetes mellitus, and hypertension. Recently, we demonstrated the ability of L-PGDS to induce apoptosis in a variety of cell types including epithelial cells, neuronal cells, and vascular smooth muscle cells (VSMCs). The aim of this study was to investigate the effect several site-directed mutations had on L-PGDS-induced apoptosis in order to identify potential sites of regulation. Point mutations created in a glycosylation site (Asn51), a protein kinase C phosphorylation site (Ser106), and the enzymatic active site (Cys65) all inhibited L-PGDS-induced apoptosis as determined by both terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) and caspase3 activity. We also compared the L-PGDS isoforms present in GK rat serum to WKY control serum using two-dimensional gel electrophoresis and observed distinct differences which vanished after PNGase F glycolytic digestion. We conclude that post-translational modification of L-PGDS, by either glycosylation or phosphorylation, enhances its apoptotic activity and inhibits VSMC hyperproliferation and postulate that this process is altered in type 2 diabetes.
- Subjects :
- Male
PNGase F
Glycosylation
Physiology
Molecular Sequence Data
Apoptosis
Biology
Biochemistry
Article
Muscle, Smooth, Vascular
chemistry.chemical_compound
Animals
Humans
Electrophoresis, Gel, Two-Dimensional
Amino Acid Sequence
Rats, Wistar
Cell Proliferation
Pharmacology
TUNEL assay
Kinase
Prostaglandin D2 synthase
Cell Biology
Molecular biology
Lipocalins
Rats
Intramolecular Oxidoreductases
Isoenzymes
Glucose
chemistry
Terminal deoxynucleotidyl transferase
Hyperglycemia
Mutation
Mutagenesis, Site-Directed
biology.protein
Phosphorylation
Protein Processing, Post-Translational
Subjects
Details
- ISSN :
- 10988823
- Volume :
- 83
- Database :
- OpenAIRE
- Journal :
- Prostaglandins & Other Lipid Mediators
- Accession number :
- edsair.doi.dedup.....45eca1f919a8c48f8e46dd4e68d390b6