Premium
Examination of isoelectric focusing and electrophoretic methods for resolving acidic proteins
Author(s) -
Green Eric D.,
Knecht David A.,
Dimond Randall L.
Publication year - 1986
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.1150070904
Subject(s) - isoelectric focusing , electrophoresis , agarose , isoelectric point , chromatography , chemistry , dictyostelium discoideum , slime mold , biochemistry , gel electrophoresis , sulfation , enzyme , gene
Highly acidic proteins are difficult to study using conventional electrofocusing and electrophoretic techniques. For example, the lysosomal enzymes from the cellular slime mold Dictyostelium discoideum have isoelectric points (p I ) around 3–4 due to extensive phosphorylation and sulfation. To assess differences in anionic modifications among these lysosomal enzymes, we required methods that resolved acidic proteins on the basis of their net charge. The separation procedures needed to be rapid and convenient, allowing parallel analysis of a large number of samples. Using these criteria, we developed two methods, isoelectric focusing in horizontal agarose gels and native electrophoresis in vertical polyacrylamide gels, which are more useful than other available techniques. Since the mechanism by which each system separates proteins is different, both methods give important information about acidic proteins.