Modification of brain glutamate dehydrogenase isoproteins with pyridoxal 5'-phosphate

被引:16
作者
Cho, SW [1 ]
Lee, JE [1 ]
机构
[1] YONSEI UNIV,COLL MED,DEPT ANAT,SEOUL 120752,SOUTH KOREA
关键词
glutamate dehydrogenase; isozymes; pyridoxal 5'-phosphate; essential lysine;
D O I
10.1016/S0300-9084(97)84333-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two soluble forms of brain glutamate dehydrogenase isoproteins were inactivated by pyridoxal 5'-phosphate. Restoration of catalytic activity can be accomplished by dialysis and addition of an excess of cysteine or lysine. Spectral evidence is presented to indicate that the inactivation proceeds through Schiff base formation with amino groups of the enzyme. Inactivation became irreversible after reduction with NaBH4 and the NaBH4-reduced enzyme showed a characteristic absorption peak at 325 nm. Using spectral titration at 325 nm, the stoichiometry was 2 mol/mol of GDH subunit without protection and 1 mol/mol with protection, indicating the complete masking of one mol of lysine. The results with analogs of pyridoxal 5'-phosphate show that the aldehyde group, but not the phosphate group, is required for efficient inactivation.
引用
收藏
页码:817 / 821
页数:5
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