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.
To determine simultaneously the relative molecular mass (M(r)) of a native oligomeric protein, and the number and M(r) of its subunits, a method using two-dimensional electrophoresis was developed. To determine the M(r) 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 M(r) of subunits. The advantage, accuracy, limitations and application of the method are discussed.
Yamaoka
T
T
Division of Endocrinology and Metabolism, University of Tsukuba, Ibaraki, Japan.
Yamashita
K
K
Itakura
M
M
D000582:Amidophosphoribosyltransferase / Q000737:chemistry*; D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D005069:Evaluation Studies as Topic; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D011487:Protein Conformation; D051381:Rats
D000582:Amidophosphoribosyltransferase