Isolation and partial characterization of two porcine spleen ferritin fractions with different electrophoretic mobility.
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.
van Gelder
W
W
Department of Chemical Pathology, Erasmus University Rotterdam, The Netherlands.
Huijskes-Heins
M I
MI
Klepper
D
D
van Noort
W L
WL
Cleton-Soeteman
M I
MI
van Eijk
H G
HG
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002847:Chromatography, Agarose; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000737:chemistry / Q000302:isolation & purification* / Q000648:ultrastructure; D006031:Glycosylation; D007120:Immunochemistry; D008854:Microscopy, Electron; D008970:Molecular Weight; D013154:Spleen / Q000737:chemistry*; D013552:Swine
D000596:Amino Acids; D005293:Ferritins