Purification and some properties of the fast migrating alkaline phosphatase in FL-amnion cells (the Kasahara isoenzyme) and its cDNA cloning.
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.
Higashino
K
K
3rd Department of Internal Medicine, Hyogo College of Medicine, Japan.
Muratani
K
K
Hada
T
T
Imanishi
H
H
Amuro
Y
Y
Yamamoto
Y
Y
Furuyama
J
J
Hirano
K
K
Hong
Y M
YM
Shimokura
M
M
D000469:Alkaline Phosphatase / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D000650:Amnion / Q000201:enzymology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000032:analysis / Q000302:isolation & purification*; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007422:Intestines / Q000201:enzymology; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010920:Placenta / Q000201:enzymology; D012333:RNA, Messenger / Q000032:analysis; D015183:Restriction Mapping
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D004274:DNA, Recombinant; D007527:Isoenzymes; D012333:RNA, Messenger; D000469:Alkaline Phosphatase