Abstract
Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. A distinct feature of IECs is that they differentiate as they migrate upwards from the crypt base to the villus tip. In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. At day 21, when cells appear to be completely differentiated, remodeling of the cell surface glycome ceases. Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract.
Highlights
From the ‡Department of Chemistry, University of California, Davis, California 95616; §Department of Dermatology, University of California, Davis School of Medicine, Sacramento, California 95816
An increase in the expression of mannosidases in partially differentiated cells is consistent with the observed decreases in high mannose type glycans and concurrent increases in complex type glycans observed in the glycomic profiles (Fig. 2)
Differentiation of Caco-2 cells grown under the conditions described here begins between days 5 and 7, with the emergence of extended microvilli structures and detectable levels of intestinal alkaline phosphatase activity
Summary
From the ‡Department of Chemistry, University of California, Davis, California 95616; §Department of Dermatology, University of California, Davis School of Medicine, Sacramento, California 95816. Precise identification of glycan compositions with structural detail and additional glycoproteomic analysis is necessary to adequately monitor changes in glycosylation patterns associated with cell differentiation. The most abundant glycans on undifferentiated cell surfaces were high mannose type structures, collectively summing to about 30% of the total signal.
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