title: Pectin of Prunus domestica L. alters sulfated structure of cell-surface heparan sulfate in differentiated Caco-2 cells through stimulation of heparan sulfate 6-O-endosulfatase-2.
abstract: Although previous reports have suggested that pectin induces morphological changes of the small intestine in vivo, the molecular mechanisms have not been elucidated. As heparan sulfate plays important roles in development of the small intestine, to verify the involvement of heparan sulfate (HS) in the pectin-induced morphological changes of the small intestine, the effects of pectin from Prunus domestica L. on cell-surface HS were investigated using differentiated Caco-2 cells. Disaccharide compositional analysis revealed that sulfated structures of HS were markedly changed by pectin administration. Real-time RT-PCR showed that pectin upregulated human HS 6-O-endosulfatase-2 (HSulf-2) expression and markedly inhibited HSulf-1 expression. Furthermore, inhibition analysis suggested that pretreatment with fibronectin III1C fragment, RGD peptide, and ERK1/2 inhibitor suppressed pectin-induced HSulf-2 expression. These observations indicate that pectin induced the expression of HSulf-2 through the interaction with fibronectin, α5β1 integrin, and ERK1/2, thereby regulating the sulfated structure of HS on differentiated Caco-2 cells.
authors.affiliation: a United Graduate School of Agricultural Science, Gifu University , Gifu , Japan.
authors.forename: Mitsutaka
authors.initials: M
authors.lastname: Nishida
authors.forename: Kazuma
authors.initials: K
authors.lastname: Murata
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kanamaru
authors.forename: Tomio
authors.initials: T
authors.lastname: Yabe
mesh_terms: D018938:Caco-2 Cells; D002454:Cell Differentiation / Q000187:drug effects*; D004187:Disaccharides / Q000032:analysis; D005353:Fibronectins / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism; D007421:Intestine, Small / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D010368:Pectins / Q000737:chemistry / Q000494:pharmacology*; D027861:Prunus / Q000737:chemistry*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013429:Sulfatases; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism*
chemical_list: D004187:Disaccharides; D005353:Fibronectins; D006497:Heparan Sulfate; D039081:Integrin alpha5beta1; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D010368:Pectins; D012333:RNA, Messenger; D013429:Sulfatases; D015238:Sulfotransferases; C470791:SULF1 protein, human; C404189:heparan sulfate 6-O-sulfotransferase; C470792:SULF2 protein, human
keywords: HSulf-2; differentiated Caco-2 cells; heparan sulfate; pectin