The posttranslational processing of sucrase-isomaltase in HT-29 cells is a function of their state of enterocytic differentiation.
The biosynthesis of sucrase-isomaltase was compared in enterocyte-like differentiated (i.e., grown in the absence of glucose) and undifferentiated (i.e., grown in the presence of glucose) HT-29 cells. Unlike differentiated cells, in which the enzyme is easily detectable and active, undifferentiated cells display almost no enzyme activity and the protein cannot be detected by means of cell surface immunofluorescence or immunodetection in membrane-enriched fractions or cell homogenates. Pulse experiments with L-[35S]-methionine show that the enzyme is, however, synthesized in these undifferentiated cells. As compared with the corresponding molecular forms in differentiated cells, the high-mannose form of the enzyme in undifferentiated cells is similarly synthesized and has the same apparent Mr. However, its complex form is less labeled and has a lower apparent Mr. Pulse-chase experiments with L-[35S]methionine show that, although the enzyme is synthesized to the same extent in both situations, the high-mannose and complex forms are rapidly degraded in undifferentiated cells, with an apparent half-life of 6 h, in contrast to differentiated cells in which the enzyme is stable for at least 48 h. A comparison of the processing of the enzyme in both situations shows that the conversion of the high-mannose to the complex form is markedly decreased in undifferentiated cells. These results indicate that the absence of sucrase-isomaltase expression in undifferentiated cells is not the consequence of an absence of biosynthesis but rather the result of both an impaired glycosylation and a rapid degradation of the enzyme.
D000911:Antibodies, Monoclonal; D002454:Cell Differentiation; D002460:Cell Line; D003110:Colonic Neoplasms; D005455:Fluorescent Antibody Technique; D006801:Humans; D007700:Kinetics; D009097:Multienzyme Complexes / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D013394:Sucrase-Isomaltase Complex / Q000096:biosynthesis / Q000235:genetics*
D000911:Antibodies, Monoclonal; D009097:Multienzyme Complexes; D013394:Sucrase-Isomaltase Complex