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Determination of molecular mass, stokes' radius, frictional coefficient and isomer‐type of non‐denatured proteins by time‐dependent pore gradient gel electrophoresis
Author(s) -
Rothe Gunter M.
Publication year - 1988
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.1150090705
Subject(s) - isocitrate dehydrogenase , chemistry , molecular mass , dehydrogenase , peroxidase , electrophoresis , malate dehydrogenase , isozyme , glutamate dehydrogenase , chromatography , esterase , gel electrophoresis , biochemistry , enzyme , glutamate receptor , receptor
Molecular mass, Stokes' radius, frictional coefficient and isomer‐type of non‐denatured proteins can be obtained by time‐dependent gradient gel electrophoresis by evaluating the resulting data using a two‐step mathematical procedure. Provided a histochemical staining procedure is available to locate the position of an enzyme in the gel, crude cell extracts can be used for estimating their molecular size properties. The computation of molecular properties of non‐denatured proteins is demonstrated for isozymes of aspartate aminotransferase (EC 2.6.1.1), peroxidase (EC 1.11.1.42) and glucose‐6‐phosphate dehydrogenase (EC 1.1.1.49) from current‐year needles of spruce. The resulting data as well as those which were calculated for esterase (EC 3.1.1.1), glutamate dehydrogenase (EC 1.4.1.4), isocitrate dehydrogenase (EC 1.1.1.42) and shikimate dehydrogenase (EC 1.1.1.25) are in accordance with those reported in the literature. The method described may be applied to various scientific areas such as genetics or environmental pollution. It could be shown here that current‐year needles of injured spruce (damage class 3) contained two more peroxidase isozymes and one more glucose‐6‐phosphate dehydrogenase isozyme than those from non‐injured trees. These differences may mark two genotypes of spruce of different susceptibilities towards present‐day air and soil pollutants.