Polyclonal immunoglobulin M: location of glycation sites.
A four step purification procedure for polyclonal human serum IgM was elaborated, including ultracentrifugation, ammonium sulfate and polyethyleneglycol precipitations and diffusion-exclusion gel chromatography. IgM was glycated in vitro both in the presence of [14C]glucose and with unlabeled glucose. Influence of incubation time up to 10 days and of glucose concentration between 10 and 60 mmol/l were studied. With 10 mmol/l glucose, a molar ratio glucose/IgM of 5.7 was attained in 10 days. Increase of glucose concentration up to 60 mmol/l led to a molar ratio of 16.0. Both basal and in vitro glycation were evaluated by 3H-labeling by gel filtration and Concanavalin A-Sepharose chromatographies. Glycation occurs mainly on the heavy chains (> 85%), particularly on the Fd region.
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Departement d'Immunologie, Immunopharmacologie et Pathologie, Faculté de Pharmacie, Université Louis Pasteur, Illkirch, France.
Gugliucci
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Stahl
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D001665:Binding Sites; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D019270:Fructosamine; D005947:Glucose / Q000378:metabolism*; D006031:Glycosylation; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000378:metabolism; D014316:Tritium; D014357:Trypsin / Q000378:metabolism
D006595:Hexosamines; D007075:Immunoglobulin M; D010446:Peptide Fragments; D014316:Tritium; D019270:Fructosamine; D014357:Trypsin; D005947:Glucose