Quantification of prostaglandin D synthetase in cerebrospinal fluid: a potential marker for brain tumor.
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.
Saso
L
L
Population Council, Rockefeller University, New York, New York 10021, USA.
Leone
M G
MG
Sorrentino
C
C
Giacomelli
S
S
Silvestrini
B
B
Grima
J
J
Li
J C
JC
Samy
E
E
Mruk
D
D
Cheng
C Y
CY
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D014408:Biomarkers, Tumor / Q000134:cerebrospinal fluid*; D001932:Brain Neoplasms / Q000134:cerebrospinal fluid* / Q000175:diagnosis; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000134:cerebrospinal fluid* / Q000737:chemistry / Q000302:isolation & purification; D054834:Lipocalins; D008969:Molecular Sequence Data; D009422:Nervous System Diseases / Q000134:cerebrospinal fluid
D014408:Biomarkers, Tumor; D054834:Lipocalins; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase