大鼠肺中CD38/ADP-核糖基环化酶的纯化与特性分析

Purification and characterization of CD38/ADP-ribosyl cyclase from rat lung.

作者信息

Khoo K M, Chang C F

机构信息

Department of Biochemistry, Faculty of Medicine, National University of Singapore, Singapore.

出版信息

Biochem Mol Biol Int. 1998 Apr;44(4):841-50. doi: 10.1080/15216549800201892.

DOI:10.1080/15216549800201892

PMID:9584998

Abstract

Purification and characterization of CD38/ADP-ribosyl cyclase in the rat lung tissue was performed with microsomes solubilized in Triton X-100 and the ADP-ribosyl cyclase was then purified using sequential column chromatography. Partially purified rat lung ADP-ribosyl cyclase was analyzed by immunoblotting using an antibody raised against a recombinant rat CD38 and showed the presence of monomer (42 kDa) and dimer (85 kDa) under non-reducing conditions but under reducing conditions, only the monomer was detected. Both the monomer and dimer could be eluted out in a stable manner from SDS-PAGE and the enzymatic activity was retained by the two different forms of CD38/ADP-ribosyl cyclase. Immunohistochemical staining showed the presence of CD38 on the bronchial epithelium and the alveoli.

摘要

采用溶解于Triton X-100的微粒体对大鼠肺组织中的CD38/ADP-核糖基环化酶进行纯化和表征，然后通过连续柱色谱法纯化ADP-核糖基环化酶。使用针对重组大鼠CD38产生的抗体通过免疫印迹分析部分纯化的大鼠肺ADP-核糖基环化酶，结果显示在非还原条件下存在单体（42 kDa）和二聚体（85 kDa），但在还原条件下，仅检测到单体。单体和二聚体均可从SDS-PAGE中稳定洗脱，且两种不同形式的CD38/ADP-核糖基环化酶均保留酶活性。免疫组织化学染色显示支气管上皮和肺泡中存在CD38。