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Characterization of human catalase by isoelectric focusing in presence of urea
Author(s) -
Alonso Antonio,
Visedo Guillemo,
Sancho Manuel,
FernándezPiqueras José
Publication year - 1990
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150110810
Subject(s) - catalase , urea , isoelectric focusing , neuraminidase , biochemistry , chemistry , sialic acid , isoelectric point , polyacrylamide gel electrophoresis , hemoglobin , enzyme , chromatography
Human catalase from erythrocytes and liver were analyzed by polyacrylamide gel isoelectric focusing in presence and absence of urea using two different pH gradients, namely pH 6–8 and pH 6.7–7.7. In presence of urea, human catalase focused in the pH range 6.75–7.0, slightly anodal to that of hemoglobin A. In narrow pH gradients, human erythrocyte catalase was microheterogeneous. Neuraminidase from different sources and peptide‐ N ‐glycosidase F were applied to investigate the presence of sialic acid and/or carbohydrate chains in human catalase. A shift in the focusing pattern of both erythrocyte and liver catalase towards the anode was observed after treatment with one of the commercially available neuraminidase preparations. This unusual result could be related to a contaminating protease since no effect was observed when the catalases were treated in presence of a serine protease inhibitor. In contrast, bovine liver and Maccaca erythrocyte catalase did not display any detectable change in their focusing patterns after treatment with any of the neuraminidase preparations.