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Different Antigenic Reactivities of Bovine Brain Glutamate Dehydrogenase Isoproteins
Author(s) -
Choi Soo Young,
Hong Joung Woo,
Song MinSun,
Jeon Seong Gyu,
Bahn Jae Hoon,
Lee Byung Ryong,
Ann JeeYin,
Cho SungWoo
Publication year - 1999
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1999.0722162.x
Subject(s) - glutamate dehydrogenase , monoclonal antibody , microbiology and biotechnology , antibody , epitope , biochemistry , enzyme , biology , chemistry , affinity chromatography , glutamate receptor , receptor , immunology
The structural differences between two types of glutamate dehydrogenase (GDH) 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 sulfatepolyacrylamide gel electrophoresis and immunoblot. Of seven anti‐GDH II monoclonal antibodies tested in the immunoblot analysis, all seven antibodies interacted with GDH II, whereas only four antibodies recognized the protein band of the other GDH isoprotein, GDH I. When inhibition tests of the GDH isoproteins were performed with the seven anti‐GDH II monoclonal antibodies, three antibodies inhibited GDH II activity, whereas only one antibody inhibited GDH i activity. The binding affinity of anti‐GDH II monoclonal antibodies for GDH II ( K D = 1.0 n M) determined using a biosensor technology (Pharmacia BIAcore) was fivefold higher than for GDH I ( K D = 5.3 n M ), These results, together with epitope mapping 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 crossreactivities of brain GDHs from some mammalian and an avian species, showing that the mammalian brain GDH enzymes are related immunologically to each other.