Reversible modification of amino groups in aspartate aminotransferase.
Amino  groups in the pyridoxal phosphate, pyridoxamine phosphate, and apo forms of pig heart cytoplasmic aspartate aminotransferase (L-aspartate: 2-oxoglutarate aminotransferase, EC .2.6.1.1) have been reversibly modified with 2,4-pentanedione. The rate of modification has been measured spectrophotometrically by observing the formation of the enamine produced and this rate has been compared with the rate of loss of catalytic activity for all three forms of the enzyme. Of the 21 amino groups per 46 500 molecular weight, approx. 16 can be modified in the pyridoxal phosphate form with less than a 50% change in the catalytic activity of the enzyme. A slow inactivation occurs which is probably due to reaction of 2,4-pentanedione with the enzyme-bound pyridoxal phosphate. The pyridoxamine phosphate enzyme is completely inactivated by reaction with 2,4-pentanedione. The inactivation of the pyridoxamine phosphate enzyme is not inhibited by substrate analogs. A single lysine residue in the apoenzyme reacts approx. 100 times faster with 2,4-pentanedione than do other amino groups. This lysine is believed to be lysine-258, which forms a Schiff base with pyridoxal phosphate in the holoenzyme.
D000588:Amines; D000818:Animals; D001051:Apoenzymes / Q000378:metabolism; D001219:Aspartate Aminotransferases / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D005977:Glutarates / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007659:Ketones / Q000494:pharmacology*; D007700:Kinetics; D008239:Lysine; D009206:Myocardium / Q000201:enzymology; D010422:Pentanones / Q000494:pharmacology*; D010710:Phosphates; D011732:Pyridoxal Phosphate; D011733:Pyridoxamine; D013056:Spectrophotometry, Ultraviolet; D013329:Structure-Activity Relationship; D013552:Swine
D000588:Amines; D001051:Apoenzymes; D005977:Glutarates; D007659:Ketones; D010422:Pentanones; D010710:Phosphates; C008790:acetylacetone; D011732:Pyridoxal Phosphate; D011733:Pyridoxamine; D001219:Aspartate Aminotransferases; D008239:Lysine