Molecular properties of rat intestinal alkaline phosphatase.
Rat intestinal alkaline phosphatase is an heterogeneous glycoprotein that contains three protein sub-forms separable by electrophoresis. The molecular weight for the glycoprotein (i.e. the average for the three sub-forms) is 157 000-160 000. Three protein sub-forms are detectable on sodium dodecyl sulphate-polyacrylamide gel electrophoresis that migrate at rates corresponding with molecular weights of 64 000, 79 000 and 92 000. Treatment of native alkaline phosphatase with 6 M guanidine - HC1 or buffer at pH 3.0 results in a product with a molecular weight of 78 000 and 70 000, respectively. Thus it is concluded that each of the three sub-forms is a dimer of identical or closely similar subunits. Limited proteolysis results in the production of new enzymically active sub-forms separable by electrophoresis. Using a bacterial protease it is possible to convert intestinal alkaline phosphatase into a form with a molecular weight of 132 000 without causing any significant change in kinetic properties. Electrophoresis of this new form on sodium dodecyl sulphate polyacrylamide gel suggests that it is composed of 66 000-dalton subunits. The native enzyme contains at least 20% by weight of carbohydrate that probably contributes to microheterogeneity of a second degree superimposed on that stemming from the presence of three protein sub-forms. Treatment with various glycosidases has no effect on electrophoretic behaviour, however. It is suggested that the three sub-forms possibly represent different stages of a maturation process that operates by limited proteolysis of a single parent protein.
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D006026:Glycoside Hydrolases; D007421:Intestine, Small / Q000201:enzymology*; D046911:Macromolecular Substances; D008970:Molecular Weight; D010447:Peptide Hydrolases; D051381:Rats
D002241:Carbohydrates; D046911:Macromolecular Substances; D000469:Alkaline Phosphatase; D006026:Glycoside Hydrolases; D010447:Peptide Hydrolases