Recombinant M-, B- and MB-type isozymes of human phosphoglyceric acid mutase: their large-scale production and preparation of polyclonal antibodies specific to M- and B-type isozymes.
Human phosphoglyceric acid mutase is a dimer comprising M-, B- and MB-type isozymes composed from the combination of the muscle-specific (M) and non-muscle-specific (B) subunits. Human DNAs coding M and B subunits were, respectively, reconstructed at their 5' regions without changing amino acid sequences, and expressed directly in Escherichia coli under the control of the trp promoter. M- and B-type isozymes were over-produced in the bacterial cytoplasm as soluble, active forms, which have been purified and characterized. MB-type was synthesized in vitro by recombining M- and B-type. All three recombinant isozymes thus obtained showed the same properties as the naturally-occurring ones with respect to the properties tested. Polyclonal IgGs specific to the M-type, B-type and MB-type were prepared from rabbits immunized with M- and B-type, using columns bound with M- and B-type. A method for the immunoassay of MB-type which is specifically present in cardiac muscle, is now under development.
Uchida
K
K
Nagahama Institute for Biochemical Science, Oriental Yeast Co., Ltd., Shiga, Japan.
Kondoh
K
K
Matuo
Y
Y
D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology; D010736:Phosphoglycerate Mutase / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis; D014169:Transformation, Bacterial
D000906:Antibodies; D007074:Immunoglobulin G; D007527:Isoenzymes; D011994:Recombinant Proteins; D004247:DNA; D010736:Phosphoglycerate Mutase