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Separation of arginase isoforms by capillary zone electrophoresis and isoelectric focusing in density gradient column
Author(s) -
Pedrosa Mercedes M.,
Legaz Maria Estrella
Publication year - 1995
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.11501601106
Subject(s) - isoelectric focusing , isoelectric point , gene isoform , chromatography , chemistry , arginase , capillary electrophoresis , electrophoresis , density gradient , biochemistry , immobilized ph gradient , gel electrophoresis , enzyme , amino acid , arginine , physics , quantum mechanics , gene
Four major arginase isoforms, I, II, III and IV, have been detected in Evernia prunastri thallus. They differ in terms of both physical and biochemical properties. The isoelectric point (p I ) of these proteins has been determined by both isoelectric focusing in density gradient column and high‐performance capillary electrophoresis (HPCE). Isoelectric focusing revealed charge microheterogeneity for isoforms II and IV whereas arginases I and II had the same p I value of 5.8. HPCE separation confirmed this charge microheterogeneity for isoform IV but not for isoform III, and provided evidence of microheterogneity for isoforms I and II. The effect of various electrolyte buffers and running conditions on the HPCE separation of arginase isoform were investigated. Addition of 0.5 mM spermidine (SPD) to the running buffer reduced the electroosmotic flow (EOF) and permitted discriminating between the native proteins and protein fragments.