Chemical modifications of histidine residues in cytoplasmic asparate aminotransferase from beef kidney
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Holo and apoenzyme of aspartate aminotransferase from beef kidney are 80% inactivated by photoxidation in the presence of 2 × 10−6m tetraiodofluroescein with the modification of two histidine residues per enzyme protomer. At a higher concentration (1 × 10−5m) a tyrosine residue is also modified. The keto substrates, ketoglutarate and oxalacetate, protect the enzyme from photoxidation.
Diethylpyrocarbonate modifies three histidine residues per enzyme protomer and reduces the activity only 10%. These results suggest that the two histidine residues photoxidized through the sensitizer, are located in the active site of the enzyme, at least one of these appears to be involved in ketosubstrate binding. The other three histidines modified by diethylpyrocarbonate are likely located on the enzyme surface and are not involved in the catalytic activity of the enzyme.
KeywordsAspartate Tyrosine Histidine Chemical Modification Catalytic Activity
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