通过高效液相色谱法对PZ肽酶活性进行高灵敏度检测。

Highly sensitive assay for PZ-peptidase activity by high-performance liquid chromatography.

作者信息

Chikuma T, Ishii Y, Kato T

出版信息

J Chromatogr. 1985 Nov 27;348(1):205-12. doi: 10.1016/s0021-9673(01)92454-9.

Abstract

A rapid and highly sensitive assay method for measuring PZ-peptidase activity in newborn rat brain is described. The method is based on monitoring the absorption at 320 nm of PZ-Pro-Leu enzymatically formed from the substrate, PZ-L-Pro-L-Leu-Gly-L-Pro-D-Arg, after separation by high-performance liquid chromatography using a reversed-phase column. This method is sensitive enough to measure PZ-Pro-Leu at concentrations as low as 5 pmol, and is able to make the column ready for the next injection within 10 min after the preceding injection. By using this method, PZ-peptidase activity was discovered in clonal osteoblastic cells derived from newborn mouse calvaria.

摘要

本文描述了一种用于测量新生大鼠脑中PZ肽酶活性的快速且高度灵敏的检测方法。该方法基于在使用反相柱进行高效液相色谱分离后，监测由底物PZ-L-Pro-L-Leu-Gly-L-Pro-D-Arg酶促形成的PZ-Pro-Leu在320nm处的吸光度。此方法灵敏度足以检测低至5pmol浓度的PZ-Pro-Leu，并且能够在前一次进样后10分钟内使色谱柱准备好进行下一次进样。通过使用该方法，在源自新生小鼠颅骨的克隆成骨细胞中发现了PZ肽酶活性。