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Quantification of prostaglandin D synthetase in cerebrospinal fluid: a potential marker for brain tumor
L Saso1, M G Leone, C Sorrentino
1Population Council, Rockefeller University, New York, New York 10021, USA.
Abstract:
Prostaglandin D synthetase (PGD-S; prostaglandin-H2 D-isomerase, EC 5,3,99,2), a 30 kDa glycoprotein also known as beta-trace protein that catalyzes the formation of prostaglandin D2 (PGD2) from PGH2, was purified to apparent homogeneity from human cerebrospinal fluid (CSF) using a two-step procedure involving HPLC on a Vydac C8 reversed-phase column and high performance electrophoresis chromatography (HPEC) using a 10% T SDS-polyacrylamide gel. The purity of PGD-S isolated from CSF was confirmed by silver stained SDS-polyacrylamide gel and direct protein microsequencing (NH2-APEAQVSVQPNFQ). A highly specific polyclonal antibody was prepared against this protein for immunoassay development. Using an ELISA, it was found that the concentration of PGD-S in CSF did not alter significantly in different pathological conditions of the central nervous system (CNS). These include dementia (n = 9), hydrocephalus (n = 4), neuropathy (n = 11), optic neuritis (n = 4), multiple sclerosis (n = 11), and demyelinating syndrome (n = 11), when compared to normal individuals (n = 12); however, the level of PGD-S in the CSF obtained from patients with brain tumor (n = 11), was reduced by as much as 2-fold when compared to control samples (n = 12) illustrating PGD-S is a potentially useful marker for brain tumor.
Insights
Prostaglandin D synthetase (PGD-S), also known as beta-trace protein, was purified from human cerebrospinal fluid. PGD-S levels were unchanged in most CNS conditions but significantly reduced in brain tumors, suggesting its potential as a diagnostic marker.
Area of Science:
- Biochemistry
- Neuroscience
- Protein Chemistry
Background:
- Prostaglandin D synthetase (PGD-S), also known as beta-trace protein, is a key enzyme in prostaglandin synthesis.
- Its role and levels in human cerebrospinal fluid (CSF) under various neurological conditions require further investigation.
Purpose of the Study:
- To purify Prostaglandin D synthetase (PGD-S) from human cerebrospinal fluid (CSF).
- To develop a specific antibody for PGD-S and establish an immunoassay for its quantification.
- To evaluate the diagnostic potential of CSF PGD-S levels in different central nervous system (CNS) pathologies.
Main Methods:
- Purification of PGD-S from human CSF using High-Performance Liquid Chromatography (HPLC) and High-Performance Electrophoresis Chromatography (HPEC).
- Confirmation of purity via SDS-polyacrylamide gel electrophoresis and protein microsequencing.
- Development of a polyclonal antibody against PGD-S for use in Enzyme-Linked Immunosorbent Assay (ELISA).
Main Results:
- PGD-S was successfully purified to homogeneity from human CSF.
- A specific polyclonal antibody was generated, enabling ELISA development.
- CSF PGD-S levels remained stable across various CNS conditions including dementia, hydrocephalus, and multiple sclerosis.
- A significant 2-fold reduction in CSF PGD-S levels was observed in patients with brain tumors compared to controls.
Conclusions:
- PGD-S can be effectively purified from human CSF.
- CSF PGD-S levels are not significantly altered in common neurological disorders.
- Reduced levels of PGD-S in CSF may serve as a potential biomarker for the early detection of brain tumors.