Influence of nucleic acids and polysaccharides on phosphotransferase activity of preparations of secretory immunoglobulin A from human milk

Biochemistry (Mosc). 2000 Feb;65(2):237-43.

Abstract

The influence of nucleic acids (DNA, tRNA), synthetic oligonucleotides, and polysaccharides (lipopolysaccharides from Escherichia coli, heparin) on protein kinase and lipid kinase activities of preparations of human secretory immunoglobulin A (sIgA) has been studied. The preparations of sIgA were isolated from human milk by chromatography on the column with Protein A-Sepharose and DEAE-sorbent (sIgA1), by affinity chromatography of sIgA1 on DNA-cellulose (sIgA2), and by gel-filtration of sIgA1 in buffer containing 5% dioxane (sIgA3). Two 32P-labeled products with high and low electrophoretic mobility in polyacrylamide gel containing SDS were found after incubation of sIgA1 and sIgA2 with [gamma-32P]ATP. The product with low electrophoretic mobility was degraded in 10% trichloroacetic acid giving a radioactive background in lanes of the polyacrylamide gel. 32P-Labeled phospholipids were found among the phosphorylation products. Soluble and immobilized DNA increase lipid kinase activity of preparations of sIgA. In this case the secretory component and H-chains of sIgA were degraded. Fractions possessing lipid kinase activity were precipitated in the presence of heparin (1 mg/ml), and lipid kinase activity was separated from sIgA by gel-filtration in buffer containing 5% dioxane. 32P-Labeled products were formed in the presence of [gamma-32P]ATP as well as [32P]ortho-phosphoric acid. The influence of heparin and synthetic deoxy- and ribooligonucleotides on casein kinase activity of sIgA3 was studied. It was observed that deoxyribooligonucleotides in micromolar concentrations increased the rate of casein phosphorylation in the presence of sIgA3 and [gamma-32P]ATP. It has been proposed that catalytically active sIgA have an affinity to DNA (anti-DNA sIgA) and can be present in human milk as a part of lipoprotein complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / pharmacology
  • Animals
  • Antibodies, Antinuclear / isolation & purification
  • Antibodies, Antinuclear / metabolism
  • Cattle
  • DNA / pharmacology
  • Female
  • Heparin / pharmacology
  • Humans
  • Immunoglobulin A, Secretory / isolation & purification
  • Immunoglobulin A, Secretory / metabolism*
  • In Vitro Techniques
  • Lipopolysaccharides / pharmacology
  • Lipoproteins / isolation & purification
  • Lipoproteins / metabolism
  • Milk Proteins / isolation & purification
  • Milk Proteins / metabolism
  • Milk, Human / enzymology*
  • Milk, Human / immunology*
  • Nucleic Acids / pharmacology
  • Phosphotransferases / isolation & purification
  • Phosphotransferases / metabolism*

Substances

  • Antibodies, Antinuclear
  • Immunoglobulin A, Secretory
  • Lipopolysaccharides
  • Lipoproteins
  • Milk Proteins
  • Nucleic Acids
  • Adenosine Triphosphate
  • Heparin
  • DNA
  • Phosphotransferases