Involvement of detergent-insoluble complexes in the intracellular transport of intestinal brush border enzymes

Biochemistry. 1995 Feb 7;34(5):1596-605. doi: 10.1021/bi00005a016.


A number of transmembrane digestive enzymes of the porcine small intestinal brush border membrane were found to be partially Triton X-100-insoluble at 0 degree C and colocalized in gradient centrifugation experiments with the GPI-anchored alkaline phosphatase in low-density, detergent-insoluble complexes commonly known as glycolipid "rafts". Thus, aminopeptidase N (EC, aminopeptidase A (EC, dipeptidyl peptidase IV (EC, and sucrase-isomaltase (EC were 34-48% detergent-insoluble. Maltase-glucoamylase (EC was markedly less detergent-insoluble (20%), and lactase-phlorizin hydrolase (EC was essentially fully soluble in detergent. In radioactively labeled, mucosal explants, the newly synthesized brush border enzymes began to associate with detergent-insoluble complexes while still in their transient, high mannose-glycosylated form, and their insolubility increased to that of the steady-state level soon after they achieved their mature, complex glycosylation, i.e., after passage through the Golgi complex. Detergent-insoluble complexes isolated by density gradient centrifugation were highly enriched in brush border enzymes, and the enrichment was apparent after only 1 h of labeling, where aminopeptidase N, sucrase-isomaltase, and alkaline phosphatase together comprised 25-30% of the total labeled, detergent-insoluble proteins, showing that sorting of newly made brush border membrane proteins into the glycolipid "rafts" does take place intracellularly. I therefore propose that, in the enterocyte, the brush border enzymes are targeted directly from the trans-Golgi network toward the apical cell surface.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminopeptidases / metabolism*
  • Animals
  • Biological Transport
  • Centrifugation, Density Gradient
  • Glycolipids / metabolism
  • Golgi Apparatus
  • Intestine, Small / metabolism*
  • Microvilli / metabolism*
  • Octoxynol / pharmacology*
  • Oligo-1,6-Glucosidase / metabolism*
  • Organ Culture Techniques
  • Swine


  • Glycolipids
  • Octoxynol
  • Oligo-1,6-Glucosidase
  • Aminopeptidases