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Isolation and partial characterization of two porcine spleen ferritin fractions with different electrophoretic mobility

By C.M. (Carin) Van Gelder, M.I.E. (M. I E) Huijskes-Heins, D. (D.) Klepper, W.L. (W.) van Noort, M.I. (Maud) Cleton-Soeteman and H.G. (Henk) van Eijk

Abstract

Ferritin isolated from porcine spleen could routinely be separated in two fractions on nondenaturating gradient gels. Both fractions could be isolated with a purity of 96% when applied to two serially linked columns, each 200 cm in length, packed respectively with Sepharose 4B and Sepharose 6B. Both fractions were similar as judged by electron microscopy. Assessed biochemically fractions were equal with respect to subunit composition, iron and phosphorus content, as well as amino acid composition (with the exception of N-acetylglucosamine). Carbohydrate analysis showed that the fraction with an apparent mass of 440 kDa (=FFL) contained 1.8% (w/w) glycans, whereas the fraction with an apparent mass of 670 kDa (=FFH) contained nearly five times as much (neutral) sugar residues (8.9%, w/w) and 10 times as much sialic acid. This difference in amount of carbohydrate side chains might explain the dissimilarity in electrophoretic mobility of the two fractions

Topics: amino acid composition, amino acid composition, carbohydrate analysis, chromatography, electron microscopy, electrophoresis, ferritin glycosylation, porcine spleen
Year: 1996
DOI identifier: 10.1016/0305-0491(96)00076-4
OAI identifier: oai:repub.eur.nl:64158

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