Corticosteroid Induced Alkaline Phosphatase: Cellular Location, Biochemical Characterization and Comparison to Intestinal Alkaline Phosphatase of the Dog (Amino Acid, Monoclonal Antibody, Peptide Mapping, Carbohydrate, N-Terminal)

The purpose of the research was to identify the subcellular location of the corticosteroid-induced isoenzyme of alkaline phosphatase (SIALP) in the liver, purify the SIALP, the intestinal isoenzyme of alkaline phosphatase (IALP) and the hepatic isoenzyme of alkaline phosphatase (LALP), develop an effective means of purification for IALP and SIALP, and characterize the purified isoenzymes. The location of the SIALP was determined to be the hepatocyte membranes which comprise the bile canaliculi of the liver. Four monoclonal antibodies against IALP were produced which cross reacted with SIALP but not with LALP. These were fused to cyanogen bromide activated sepharose and used to develop a rapid purification procedure which dramatically increased the yield of IALP and SIALP over the conventional chromatography techniques used to purify LALP. Results of amino acid analysis, peptide mapping, N-terminal sequencing of the first 10 amino acids, and carbohydrate analysis indicates SIALP and IALP are identical to each other except in carbohydrate content, and that the SIALP is either a product of the IALP gene or a product of addition of carbohydrate including sialic acid to IALP which was taken up into the hepatocyte by endocytosis.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Veterinary Medical Science

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

1986

Author dc:creator

• Sanecki, Robin Kenneth

• Biology, Veterinary Science

Identifier

(UMI)AAI8623402

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/71338