N-glycans stabilize human erythropoietin through hydrophobic interactions with the hydrophobic protein surface: studies by surface plasmon resonance analysis.
Human erythropoietin (EPO) produced in Chinese hamster ovary cells is a hydrophobic protein highly stabilized by multibranched complex-type N-glycans. To reveal the molecular basis of the interaction between the N-glycans and the EPO protein, complex-type N-glycans of different structures were analyzed as to their binding affinity for Escherichia coli-expressed EPO by means of the surface plasmon resonance technique. It appears well established that complex-type N-glycans, particularly multibranched ones, have hydrophobic regions that extensively stretch across the plane holding acetylamino groups and that N-glycan-protein hydrophobic interactions characterized by a slow rate of dissociation stabilize the protein conformation.
Toyoda
Teruko
T
Course of Applied Biochemistry, Graduate School of Agriculture and Biological Science, Osaka Prefecture University, Sakai, 599-8531, Japan.
Arakawa
Tsutomu
T
Yamaguchi
Haruki
H
D000818:Animals; D016466:CHO Cells; D002850:Chromatography, Gel; D006224:Cricetinae; D004921:Erythropoietin / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008956:Models, Chemical; D008958:Models, Molecular; D011134:Polysaccharides / Q000502:physiology*; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D020349:Surface Plasmon Resonance; D013997:Time Factors
D011134:Polysaccharides; D004921:Erythropoietin