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Qualitative Analysis of the Carbohydrate Composition of Apolipoprotein H
Authors:Roberto Gambino   Gianluca Ruiu   Gianfranco Pagano  Maurizio Cassader
Affiliation:(1) Dipartimento di Medicina Interna, Universita' di Torino, 10126 Torino, Italy;(2) Laboratorio Centrale "ldquo"Baldi e Riberi"rdquo", Azienda Ospedaliera S. Giovanni Battista, Torino, Italy
Abstract:The specific binding of digoxigenin-labeled lectins to carbohydrate moieties is used to characterize the carbohydrate chains bound to apolipoprotein H. Our results show that apolipoprotein H is rich in sialic acid linked agr(2–6) to galactose or N-acetylgalactosamine. Sialic acid is not agr(2–3)-linked to galactose. Galactose is beta(1–4)-linked to N-acetylglucosamine and beta(1–3)-linked to N-acetylgalactosamine. High-mannose N-glycan chains are barely detectable. After N-glycosidase F treatment the molecular weight is substantially reduced. The main band is 32,500 daltons. Carbohydrate O-linked chains, which are mainly represented by sialic acid, are agr(2–6)-linked to galactose or N-acetylgalactosamine. Galactose is also organized in O-linked chains and it is beta(1–4)-linked to N-acetylglucosamine and beta(1–3)-linked to acetylgalactosamine. Biochemical analysis of carbohydrate structures reveals that no specific carbohydrate complex is bound to a single isoform.
Keywords:Apolipoprotein H    /content/w6871747j567vm06/xxlarge946.gif"   alt="  beta"   align="  MIDDLE"   BORDER="  0"  >2-glycoprotein I  lectin affinity chromatography  glycosylation
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