Purification and comparison of corticosteroid-induced and intestinal isoenzymes of alkaline phosphatase in dogs

Sanecki, R.K.; Hoffmann, W.E.; Dorner, J.L.; Kuhlenschmidt, M.S.

Purification and comparison of corticosteroid-induced and intestinal isoenzymes of alkaline phosphatase in dogs

1990

Sanecki, R.K. | Hoffmann, W.E. | Dorner, J.L. | Kuhlenschmidt, M.S.

Corticosteroid-induced alkaline phosphatase (CALP) and intestinal alkaline phosphatase (IALP) from dogs were purified to homogeneity, as determined by polyacrylamide gel electrophoresis. Purification involved an uninterrupted system using DEAE-cellulose, concanavalin A-agarose, and monoclonal antibody affinity columns. The monoclonal antibody was prepared by use of IALP as the antigen. The 2 isoenzymes were compared, using molecular weight determinations, amino acid analyses, peptide mapping, N-terminal sequencing of the first 10 amino acids, carbohydrate analyses, and recognition by anti-IALP monoclonal antibody. The data indicated that canine IALP and CALP are identical with regard to recognition by monoclonal antibody and N-terminal amino acid sequence, nearly identical in amino acid content and peptide maps, but different in carbohydrate content. It was concluded that CALP is a product of the same gene as IALP and that differences in glycosyl transferase activities between liver and intestines or the presence of glycosidase activities in or around the intestinal mucosae result in the marked difference in carbohydrate content.

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