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脑脊液中前列腺素D合成酶的定量分析:一种潜在的脑肿瘤标志物。

Quantification of prostaglandin D synthetase in cerebrospinal fluid: a potential marker for brain tumor.

作者信息

Saso L, Leone M G, Sorrentino C, Giacomelli S, Silvestrini B, Grima J, Li J C, Samy E, Mruk D, Cheng C Y

机构信息

Population Council, Rockefeller University, New York, New York 10021, USA.

出版信息

Biochem Mol Biol Int. 1998 Nov;46(4):643-56. doi: 10.1080/15216549800204172.

DOI:10.1080/15216549800204172
PMID:9844724
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.

摘要

前列腺素 D 合成酶(PGD-S;前列腺素-H2 D-异构酶,EC 5.3.99.2)是一种 30 kDa 的糖蛋白,也被称为β-微量蛋白,它催化从 PGH2 形成前列腺素 D2(PGD2)。使用两步法从人脑脊液(CSF)中纯化该酶至表观均一性,第一步是在 Vydac C8 反相柱上进行高效液相色谱(HPLC),第二步是使用 10% T SDS-聚丙烯酰胺凝胶进行高效电泳色谱(HPEC)。通过银染 SDS-聚丙烯酰胺凝胶和直接蛋白质微测序(NH2-APEAQVSVQPNFQ)确认了从 CSF 中分离的 PGD-S 的纯度。制备了针对该蛋白的高特异性多克隆抗体用于免疫分析开发。使用酶联免疫吸附测定(ELISA)发现,在中枢神经系统(CNS)的不同病理状况下,CSF 中 PGD-S 的浓度没有显著变化。这些病理状况包括痴呆(n = 9)、脑积水(n = 4)、神经病变(n = 11)、视神经炎(n = 4)、多发性硬化症(n = 11)和脱髓鞘综合征(n = 11),与正常个体(n = 12)相比;然而,与对照样本(n = 12)相比,从脑肿瘤患者(n = 11)获得的 CSF 中 PGD-S 的水平降低了多达 2 倍,说明 PGD-S 是脑肿瘤的一个潜在有用标志物。

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