Different antigenic reactivities of bovine brain glutamate dehydrogenase isoproteins

The structural differences between two types of glutamate dehydrogenase (GD H) isoproteins (GDH I and GDH II), homogeneously isolated from bovine brain , were investigated using a biosensor technology and monoclonal antibodies. A total of seven monoclonal antibodies raised against GDH II were produced , and the antibodies recognized a single protein band that comigrates with purified GDH II on sodium dodecyl sulfate-polyacrylamide gel electrophoresi s and immunoblot, Of seven anti-GDH II monoclonal antibodies tested in the immunoblot analysis, all seven antibodies interacted with GDH II, whereas o nly four antibodies recognized the protein band of the other GDH isoprotein , GDH I, When inhibition tests of the GDH isoproteins were performed with t he seven anti-GDH II monoclonal antibodies, three antibodies inhibited GDH II activity, whereas only one antibody inhibited GDH I activity. The bindin g affinity of anti-GDH II monoclonal antibodies for GDH II (K-D = 1.0 nM) d etermined using a biosensor technology (Pharmacia BIAcore) was fivefold hig her than for GDH I (K-D = 5.3 nM), These results, together with epitope map ping analysis, suggest that there may be structural differences between the two GDH isoproteins, in addition to their different biochemical properties . Using the anti-GDH II antibodies as probes, we also investigated the cros sreactivities of brain GDHs from some mammalian and an avian species, showi ng that the mammalian brain GDH enzymes are related immunologically to each other.