title: Characterisation of rat and human tissue alkaline phosphatase isoforms by high-performance liquid chromatography and agarose gel electrophoresis.
abstract: Alkaline phosphatase (ALP) exists as several isoenzymes and many isoforms present in tissues and serum. The objective of this study was to separate tissue ALP forms in rats and humans and characterise their properties. The materials for the investigation were intestinal, bone, and liver tissue of rats and commercially available human preparations of tissue ALP. Two methods of separation were used: high-performance liquid chromatography (HPLC) and agarose gel electrophoresis. Using HPLC in the rat tissues, two ALP isoforms in the intestine, one in the bone, and three in the liver were identified. In humans three intestinal, two bone, and one liver isoform were resolved. Electrophoresis showed two ALP activity bands in rat intestine, one wide band in the bone, and three bands in the liver. ALP of human tissues was visualised as a single wide band, with a different mobility observed for each organ. In both species the presence of a form with properties characteristic of the bone isoform of the tissue-nonspecific isoenzyme was observed in the intestine. HPLC offers a higher resolution than electrophoresis with respect to tissue ALP fractions in rats and in humans, but electrophoresis visualises high-molecular-mass insoluble enzyme forms.
authors.affiliation: Department of Biochemistry and Medical Chemistry, Pomeranian Medical University, Al. Powstańców Wlkp. 72, 70-111 Szczecin, Poland. viola@sci.pam.szczecin.pl
authors.forename: Violetta
authors.initials: V
authors.lastname: Dziedziejko
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Safranow
authors.forename: Dorota
authors.initials: D
authors.lastname: Slowik-Zylka
authors.forename: Anna
authors.initials: A
authors.lastname: Machoy-Mokrzynska
authors.forename: Barbara
authors.initials: B
authors.lastname: Millo
authors.forename: Zygmunt
authors.initials: Z
authors.lastname: Machoy
authors.forename: Dariusz
authors.initials: D
authors.lastname: Chlubek
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis*; D000818:Animals; D001409:Bacillus cereus / Q000201:enzymology; D001842:Bone and Bones / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D054800:Glycosylphosphatidylinositol Diacylglycerol-Lyase / Q000378:metabolism* / Q000494:pharmacology; D006358:Hot Temperature; D006801:Humans; D007422:Intestines / Q000201:enzymology; D007527:Isoenzymes / Q000032:analysis*; D007700:Kinetics; D037102:Lectins / Q000494:pharmacology; D008099:Liver / Q000201:enzymology; D008297:Male; D010649:Phenylalanine / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D014508:Urea / Q000494:pharmacology
chemical_list: D007527:Isoenzymes; D037102:Lectins; D010649:Phenylalanine; D014508:Urea; D000469:Alkaline Phosphatase; D054800:Glycosylphosphatidylinositol Diacylglycerol-Lyase