Posttranslational regulation of sucrase-isomaltase expression in intestinal crypt and villus cells

J Biol Chem. 1989 Nov 25;264(33):20000-11.


Expression and synthesis of sucrase-isomaltase (SI) were studied in human jejunum and in the colon tumor cell lines Caco-2 and HT-29. Twelve monoclonal antibodies produced against the adult human intestinal enzyme were shown to recognize specifically SI by immunoprecipitation of 14C-labeled membrane proteins, analysis of enzyme activities in the immunoprecipitates, and immunoblotting. These antibodies produced markedly different patterns of immunofluorescent staining of the intestinal mucosa. Three of them were specific for the absorptive villus cells, while the other nine also stained the luminal membrane of the proliferative crypt cells, with different intensities which paralleled their ability to recognize SI in immunoblots. Sequential immunoprecipitation of SI solubilized from purified brush borders or entire jejunum with four selected antibodies demonstrated the presence of different forms of the enzyme, expressed by either villus or crypt cells. Two immunologically distinct forms of high mannose precursor (hmP1 and hmP2) were also identified in both jejunal mucosa and colon tumor cells. They were present as monomers and their immunological differences were preserved under various ionic and pH conditions. Pulse-chase studies indicated that, in Caco-2 cells, hmP1 is converted into hmP2 within 30 min of chase, and hmP2 is then processed into the complex-glycosylated precursor destined for the brush border membrane. hmP1 was immunologically related to the mature SI present in crypt cells and lacked the epitopes specific for mature SI expressed by villus cells. These results demonstrated that sucrase-isomaltase is synthesized by both crypt and villus cells, but processing of the cotranslationally glycosylated high mannose precursor is dependent on the state of differentiation of the enterocytes. This may represent a general mechanism for the regulation of expression of differentiated cell products at the post-translational level.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Animals
  • Antibodies, Monoclonal
  • Electrophoresis, Gel, Two-Dimensional
  • Electrophoresis, Polyacrylamide Gel
  • Epithelium / enzymology
  • Fluorescent Antibody Technique
  • Gene Expression*
  • Humans
  • Immunoblotting
  • Intestinal Mucosa / enzymology*
  • Jejunum / enzymology
  • Kinetics
  • Macromolecular Substances
  • Mice
  • Mice, Inbred BALB C
  • Microvilli / enzymology*
  • Multienzyme Complexes / genetics*
  • Protein Processing, Post-Translational*
  • Sucrase-Isomaltase Complex / analysis
  • Sucrase-Isomaltase Complex / genetics*
  • Sucrase-Isomaltase Complex / metabolism


  • Antibodies, Monoclonal
  • Macromolecular Substances
  • Multienzyme Complexes
  • Sucrase-Isomaltase Complex