title: Multiple forms of human intestinal alkaline phosphatase: chemical and enzymatic properties, and circulating clearances of the fast- and slow-moving enzymes.
abstract: Two forms of alkaline phosphatase (orthophosphoric monoester phosphohydrolase (alkaline optimum, EC 3.1.3.1) have been purified from human small intestine by column chromatography on DEAE-cellulose and tyraminyl derivative affinity gel, and by preparative disc gel electrophoresis. Intestinal phosphatases were electrophoretically separated into two components, fast- and slow-moving enzymes, with apparent molecular weights of 140000 and 168000 and with subunit weights of 68000 and 80000, respectively. Analyses of carbohydrate and amino acid revealed marked differences in the two enzymes. Enzymatic properties and affinities for an anti-blood group antibody were also found to differ. Papain digestion released a hydrophobic small peptide from the slow-moving enzyme and its enzymatic properties resembled those of the fast-moving enzyme. Circulating clearance (T1/2) of the slow- and fast-moving enzymes from adult intestine was found to be 7.5 h and 1.3 h, respectively; that of fetal intestinal enzyme was 2.8 h. Sialidase, sialidase/beta-galactosidase, or sialidase/beta-galactosidase/N-acetyl-beta-glucosaminidase treatment of the fetal enzyme reduced the value to about 40 min. Further, digestion with alpha-fucosidase, alpha-mannosidase or both restored it to nearly the original level. Organ distribution of injected 125I-labelled enzymes indicates that the desialylated hepatic enzyme was selectively distributed in liver, while the degalactosylated intestinal enzyme was incorporated into liver lymph fluid, and small intestine. These results suggest that the pathway of circulating clearance of alkaline phosphatase has several routes.
mesh_terms: D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis*; D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D005260:Female; D006026:Glycoside Hydrolases; D006801:Humans; D007231:Infant, Newborn; D007422:Intestines / Q000201:enzymology*; D007527:Isoenzymes / Q000032:analysis; D008670:Metals / Q000032:analysis; D008970:Molecular Weight; D010206:Papain; D011247:Pregnancy; D011506:Proteins / Q000032:analysis
chemical_list: D000596:Amino Acids; D002241:Carbohydrates; D007527:Isoenzymes; D008670:Metals; D011506:Proteins; D000469:Alkaline Phosphatase; D006026:Glycoside Hydrolases; D010206:Papain