Nyberg F, Terenius L
Neuropeptides. 1985 Feb;5(4-6):537-40. doi: 10.1016/0143-4179(85)90073-3.
An enzyme radioimmunoassay procedure has been used to identify (Leu)- and (Met)enkephalin containing polypeptides in human cerebrospinal fluid (CSF). A large volume of lumbar CSF collected from 15-20 neurologic subjects was fractionated by ion exchange chromatography on Aminex Q-15 and subsequently on Sephadex G-50. The lyophilized fractions were treated with trypsin and the generated enkephalyl hexapeptides were isolated by SP-Sephadex ion-exchange chromatography and quantified by radioimmunoassay specific for (Met)enkephalin-Arg6/Lys6 and (Leu)enkephalin-Arg6, respectively. Confirmation of the structures of the recovered fragments was obtained by HPLC. In total, the procedure yielded about 100 fmole (Met)-enkephalyl and 40 fmole (Leu)enkephalyl hexapeptides per ml CSF. Our results also suggest that the main part of the (Leu)enkephalin related activity emerged from prodynorphin.
已采用酶放射免疫分析方法来鉴定人脑脊液（CSF）中含亮氨酸脑啡肽和甲硫氨酸脑啡肽的多肽。从15至20名神经科患者收集的大量腰椎脑脊液通过在Aminex Q - 15上进行离子交换色谱分离，随后在Sephadex G - 50上进行分离。冻干的组分用胰蛋白酶处理，产生的脑啡肽六肽通过SP - Sephadex离子交换色谱分离，并分别通过对甲硫氨酸脑啡肽 - Arg6/Lys6和亮氨酸脑啡肽 - Arg6特异的放射免疫分析进行定量。通过高效液相色谱法对回收片段的结构进行了确认。总体而言，该方法每毫升脑脊液产生约100飞摩尔的甲硫氨酸脑啡肽六肽和40飞摩尔的亮氨酸脑啡肽六肽。我们的结果还表明，与亮氨酸脑啡肽相关的活性主要来自前强啡肽。
