Purification and partial characterization of intestinal-like alkaline phosphatase in rabbit kidney.
Two types of alkaline phosphatase (AP) isozymes in rabbit kidney, a major intestinal-like type and a minor tissue-unspecific type, have been identified. The former enzyme was purified from rabbit kidney by immunoaffinity chromatography using monoclonal anti-human intestinal AP antibody. The purified enzyme yielded a single protein band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the apparent molecular size of its monomer subunit was found to be 72,000. Three amino acid residues within the first 16 N-terminal amino acid residues were different in purified AP and human intestinal AP. Although the rabbit enzyme possessed some peptide bands identical to those of human adult intestinal AP after Staphylococcus aureus V8 protease digestion, the enzyme did not react with monoclonal antibody against human adult intestinal AP alone, whereas it did react with monoclonal antibody against both human adult and fetal intestinal APs. The affinity of the enzyme for concanavalin A was identical to that of the fetal intestinal AP, but different from that of the adult enzyme. These results indicate that the antigenicity and certain properties of purified rabbit AP are more like those of human fetal intestinal AP or Kasahara isozyme, so-called intestinal-like AP, than like human adult intestinal AP.
Fujimori-Arai
Y
Y
Department of Biochemistry, Saitama Medical School, Japan.
Koyama
I
I
Hirano
K
K
Sakagishi
Y
Y
Komoda
T
T
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002241:Carbohydrates / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006868:Hydrolysis; D007422:Intestines / Q000187:drug effects / Q000201:enzymology*; D007527:Isoenzymes / Q000276:immunology; D007668:Kidney / Q000187:drug effects / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D012697:Serine Endopeptidases / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D007527:Isoenzymes; D000469:Alkaline Phosphatase; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase