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Determination of the number and relative molecular mass of subunits in an oligomeric protein by two-dimensional electrophoresis Application to the subunit structure analysis of rat liver amidophosphoribosyltransferase |
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| Authors: | Takashi Yamaoka and Kamejiro Yamashita Mitsuo Itakura |
| Institution: | Division of Endocrinology and Metabolism, Institute of Clinical Medicine, University of Tsukuba, Tsukuba-City, Ibaraki 305 Japan Otsuka Department of Clinical and Molecular Nutrition, University of Tokushima, School of Medicine, Kuramoto-cho, Tokushima-City 770 Japan |
| Abstract: | To determine simultaneously the relative molecular mass (Mr) of a native oligomeric protein, and the number and Mr of its subunits, a method using two-dimensional electrophoresis was developed. To determine the Mr of a native oligomeric protein, pore gradient gel electrophoresis was performed for the first dimension. Native proteins were dissociated into their subunits by sodium dodecyl sulphate (SDS) in a gel slice, then applied to SDS polyacrylamide gel electrophoresis for the second dimension to determine the Mr of subunits. The advantage, accuracy, limitations and application of the method are discussed. |
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