Qualitative Analysis of the Carbohydrate Composition of Apolipoprotein H |
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Authors: | Roberto Gambino Gianluca Ruiu Gianfranco Pagano Maurizio Cassader |
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Affiliation: | (1) Dipartimento di Medicina Interna, Universita' di Torino, 10126 Torino, Italy;(2) Laboratorio Centrale Baldi e Riberi, Azienda Ospedaliera S. Giovanni Battista, Torino, Italy |
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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 (2–6) to galactose or N-acetylgalactosamine. Sialic acid is not (2–3)-linked to galactose. Galactose is (1–4)-linked to N-acetylglucosamine and (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 (2–6)-linked to galactose or N-acetylgalactosamine. Galactose is also organized in O-linked chains and it is (1–4)-linked to N-acetylglucosamine and (1–3)-linked to acetylgalactosamine. Biochemical analysis of carbohydrate structures reveals that no specific carbohydrate complex is bound to a single isoform. |
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Keywords: | Apolipoprotein H /content/w6871747j567vm06/xxlarge946.gif" alt=" beta" align=" MIDDLE" BORDER=" 0" >2-glycoprotein I lectin affinity chromatography glycosylation |
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