Serum leucine-rich alpha-2-glycoprotein-1 binds cytochrome c and inhibits antibody detection of this apoptotic marker in enzyme-linked immunosorbent assay
Serum leucine-rich alpha-2-glycoprotein-1 binds cytochrome c and inhibits antibody detection of this apoptotic marker in enzyme-linked immunosorbent assay
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DOI：
10.1007/s10495-006-8159-3
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发表时间：
2006-07-01
期刊：
影响因子：
7.2
通讯作者：
Jemmerson, Ronald
中科院分区：
文献类型：
--
作者：
Cummings, Chad;Walder, Jennifer;Jemmerson, Ronald
细胞色素 c (Cyt c) 已被认为是异常细胞凋亡的血清标志物，因此具有相当大的临床潜力。使用夹心酶联免疫吸附测定 (ELISA)，我们发现在血清存在的情况下，Cyt c 检测的灵敏度会降低。采用标准层析程序，然后在 Affi-Gel 10 结合的 Cyt c 上进行亲和层析，从胎牛血清和人血清中纯化出负责的抑制因子。在 SDS-PAGE 中，分别观察到牛蛋白和人蛋白的 44 kD 和 50 kD 条带。质谱分析确定血清抑制因子为富含亮氨酸的α-2-糖蛋白-1 (LR α 2GP1)。这种鉴定可能会导致改进的 ELISA 来量化患者血清中的总 Cyt c。 LR α 2GP1 是第一个已被鉴定的 Cyt c 细胞外配体。提出了与结合相关的生理功能。
Cytochrome c (Cyt c) has been implicated as a serum marker for aberrant apoptosis and, thus, has considerable clinical potential. Using a sandwich enzyme-linked immunosorbent assay (ELISA) we found that the sensitivity of Cyt c detection is reduced in the presence of serum. The inhibitory factor responsible was purified from both fetal bovine serum and human serum employing standard chromatography procedures followed by affinity chromatography on Affi-Gel 10-bound Cyt c. In SDS-PAGE, bands at 44 kD and 50 kD were observed for the bovine and human proteins, respectively. Mass spectrometry analysis identified the serum inhibitory factor as leucine-rich alpha-2-glycoprotein-1 (LR alpha 2GP1). This identification may lead to a modified ELISA to quantify total Cyt c in patients' sera. LR alpha 2GP1 is the first extracellular ligand for Cyt c that has been identified. A physiological function associated with binding is suggested.