Peptide hydrolases of the human small intestinal mucosa: distribution of activities between brush border membranes and cytosol

Abstract

Brush border membranes from frozen human small intestine have been purified using a method which did not involve the use of EDTA-containing buffers or the disruption of brush border fragments with high concentrations of Tris. On average a 24-fold increase in specific activity of alpha-glucosidase (brush border marker) was obtained in the final preparation which contained insignificant traces of enzyme marker activities from cytosol and lysosomes. The homogenates of human small intestinal mucosa were shown to contain enzymes capable of hydrolysing di-, tri-, and tetrapeptides as well as amino acid- and peptide-2-nephthylamides. Assuming a 100% location of alpha-glucosidase in the brush border membrane, distribution studies indicated that activities against tetrapeptides and leucyl-2-naphthylamide were located exclusively in the brush border membrane. A large proportion of activity against alpha-glutamyl-2-naphthylamide, gamma-glutamyl-2-naphthylamide and glycyl-prolyl-2-naphthylamide were also recovered in the brush border membrane fraction. Depending on the substrate utilized, 33-87% of tripeptidase activity was located in the brush border membrane. However, 58-87% of dipeptidase activity was recovered in the soluble fraction.