title: Structural studies on sugar chains of carbohydrate-deficient transferrin from patients with alcoholic liver disease using lectin affinity electrophoresis.
abstract: It is well-known that microheterogeneity of human serum transferrin observed in alcoholics manifests as sialic acid-deficient transferrin isoforms, otherwise known as carbohydrate-deficient transferrin (CDT). A recent study demonstrated that serum CDT lacked one or both of the entire carbohydrate chains but the investigation required several troublesome procedures. The aim of the present study was to confirm the sugar chain structures of serum transferrin, and of serum CDT in particular, from patients with alcoholic liver disease (ALD) using conventional lectin affinity electrophoresis which might be useful in the clinical setting. The serum CDT obtained from ALD-patients was partially purified using an anion exchanger. Serum transferrin and the partially purified serum CDT were investigated by concanavalin A (Con A)- and Datura stramonium agglutinin (DSA)-affinity electrophoresis followed by antibody-affinity blotting and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with Western blotting. By Con A-affinity electrophoresis, serum CDT was separated into weakly reactive and nonreactive transferrins which showed slower electrophoretic mobilities than those from the healthy controls. Moreover, nearly all of the serum CDT was nonreactive with DSA. On SDS-PAGE, the molecular masses of serum CDT were estimated to be approximately 75 and 72 kDa, which corresponded to those of partially and completely deglycosylated transferrin obtained from the healthy controls (78 kDa), respectively. In conclusion, these results indicated that the sugar chain structures of serum CDT from patients with ALD show not merely a loss of terminal sialic acids, but also the absence of asparagine-N-linked oligosaccharides.
authors.affiliation: Department of Internal Medicine (I), The Jikei University School of Medicine, Tokyo, Japan. ramses@mxr.meshnet.or.jp
authors.forename: T
authors.initials: T
authors.lastname: Inoue
authors.forename: M
authors.initials: M
authors.lastname: Yamauchi
authors.forename: K
authors.initials: K
authors.lastname: Ohkawa
mesh_terms: D000915:Antibody Affinity; D015153:Blotting, Western; D003208:Concanavalin A; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006801:Humans; D037102:Lectins; D008108:Liver Diseases, Alcoholic / Q000097:blood*; D019158:N-Acetylneuraminic Acid; D037121:Plant Lectins; D011487:Protein Conformation; D014168:Transferrin / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: C041384:Datura stramonium lectin; D037102:Lectins; D037121:Plant Lectins; D014168:Transferrin; C056759:carbohydrate-deficient transferrin; D003208:Concanavalin A; D019158:N-Acetylneuraminic Acid