ABSTRACT
In this study we isolated a highly purified, homogeneous high molecular weight (HMW) AcPase (Mr 102 kDa) from the chicken liver. This enzyme was shown to be a slightly acidic (pI 5.0-6.1), dimeric sialoglycoenzyme, composed of two equivalent subunits. Its sugar moiety, characterized by interactions with specific lectins, was shown to be composed of hybrid and complex type of carbohydrate chains. Heterogeneity of the high molecular weight AcPase arising from variations of the sugar components was demonstrated by isoelectric focusing, followed by reactions of the isoelectric components with specific lectins on NC membranes. Structural relationship based on immunological similarities was shown between the HMW AcPases from carp, frog, and chicken livers.
Subject(s)
Acid Phosphatase/chemistry , Chickens , Glycoproteins/chemistry , Liver/enzymology , Acid Phosphatase/immunology , Acid Phosphatase/isolation & purification , Animals , Antigens/immunology , Anura , Carps , Dimerization , Immunoblotting , Immunoglobulin G/immunology , Isoelectric Focusing , Isoelectric Point , Molecular Weight , Species SpecificityABSTRACT
The role of the carbohydrate moiety of Tamm-Horsfall protein (THP) was investigated by studying the effect of partial enzymatic desialylation and deglycosylation on its immunoreactivity. In our experiments primarily sialic acid alpha 2,6 linked to a galactose was split off, resulting in a 14% increase of immunoreactivity. It was increased up to 21% by further removal of the sugar residues. We also found that complexes composed of THP and human serum albumin, immunoglobulin G or transferrin reacted with anti-THP antibodies. When these complexes were formed with desialylated or deglycosylated THP an essential increase of this reactivity (about 25%) occurred. Our studies indicate that modifications of the carbohydrate moiety of THP (which might occur in vivo by changes in the glycosylation process e.g. in pathologic conditions) lead to increased exposure of THP to the immune system.
