We have previously characterized the expression of the alpha nu beta 5 and alpha nu beta 6 integrins as major receptors for the human colonic adenocarcinoma cell line (HT29-D4), on vitronectin and fibronectin, respectively [Lehmann et al. (1994): Cancer Res 54:2102-2107]. In the present work we investigated the glycosylation role of these integrins in their adhesive functions. To this end, we used glycohydrolases to show that cell surface integrins were N-glycosylated and sialylated, and that only the alpha v subunit carried some immature oligosaccharide side chains. To alter the glycosylation state of the cell surface alpha v beta 5 and alpha v beta 6 integrins, we used two oligosaccharide-processing inhibitors: 1-deoxymannojirimycin (dMNJ) and tunicamycin (TM). Following treatment of HT29-D4 cells with dMNJ, cell surface alpha v beta 5 and alpha v beta 6 carried only high-mannose-type sugar chains, while TM-treated cells expressed de-N-glycosylated integrins. Neither alpha/beta heterodimers assembly nor cell surface expression were impaired in the presence of the drugs. Finally, we established that adhesion of dMNJ- or TM-treated cells was altered on both vitronectin and fibronectin substrata, whereas the adhesion of these cells on laminin or collagen type I was virtually unchanged.