FL-羊膜细胞中快速迁移碱性磷酸酶（笠原同工酶）的纯化、某些性质及其cDNA克隆

Purification and some properties of the fast migrating alkaline phosphatase in FL-amnion cells (the Kasahara isoenzyme) and its cDNA cloning.

作者信息

Higashino K, Muratani K, Hada T, Imanishi H, Amuro Y, Yamamoto Y, Furuyama J, Hirano K, Hong Y M, Shimokura M

机构信息

3rd Department of Internal Medicine, Hyogo College of Medicine, Japan.

出版信息

Clin Chim Acta. 1990 Jan 15;186(2):151-64. doi: 10.1016/0009-8981(90)90032-n.

DOI:10.1016/0009-8981(90)90032-n

PMID:2311250

Abstract

One of two main FL-amnion cell alkaline phosphatase (AP), the fast migrating one (FL-APF) has been reported to be identical to Kasahara isoenzyme (K.I.), which occurs preferentially in sera of patients with primary hepatoma. We purified FL-APF of which the apparent molecular weight was 135,000 by gel filtration, and that of the subunit was 62,000 on SDS/PAGE, indicating homodimeric structure of FL-AL-APF. FL-APF was found to react with monoclonal antibody against adult intestinal AP, but not with monoclonal antibody to placental AP. We isolated FL-APF cDNA clone from FL-amnion cells, of which cDNA was 2525 base pairs in length. Nucleotide sequence of the coding region and the 3' untranslated region was identical to the sequence of human adult intestinal AP cDNA. But the untranslated region of the 5' end of the isolated clone was slightly longer than that of intestinal AP. Hence, FL-APF (K.I.) may occur by altered glycosylation of intestinal AP.

摘要

胎膜羊膜细胞碱性磷酸酶（AP）主要有两种，其中迁移速度较快的一种（FL-APF）据报道与笠原同工酶（K.I.）相同，后者优先出现在原发性肝癌患者的血清中。我们通过凝胶过滤纯化了表观分子量为135,000的FL-APF，在SDS/PAGE上其亚基分子量为62,000，表明FL-AL-APF具有同二聚体结构。发现FL-APF与抗成人肠道AP的单克隆抗体反应，但不与胎盘AP的单克隆抗体反应。我们从胎膜羊膜细胞中分离出FL-APF cDNA克隆，其cDNA长度为2525个碱基对。编码区和3'非翻译区的核苷酸序列与人成人肠道AP cDNA的序列相同。但分离克隆5'端的非翻译区比肠道AP的略长。因此，FL-APF（K.I.）可能是由肠道AP糖基化改变而产生的。