[Photooxidation of aspartate transaminase from chicken heart cytosol].
Rose-bengal-sensitized photooxidation of aspartate transaminase from chicken heart cytosol results in a loss of enzymatic activity which follow first order kinetics down to 70--75% inactivation. 0.9 Histidine, 0.9 tryptophane residues and 1.5 SH groups per enzyme subunit were found to be modified in the photooxidized transaminase, which retained 26% residual activity. Photodestruction of the coenzyme was about 16%. The rate of enzyme photoinactivation is constant in the pH range 6--8, and drastically decreases with lowering pH from 6 to 4. alpha-Ketoglutarate partially protects the holoenzyme from inactivation. The apoenzyme undergoes photoinactivation at a rate almost twice as rapid as the holoenzyme. Photooxidized apotransaminase retains affinity to pyridoxal phosphate and binds as much coenzyme as the native apoenzyme. Photooxidation induces no significant alterations in the circular dichroism pattern of the enzyme in the 200 to 240 nm range. However, positive circular dichroism is markedly increased in the absorption bands of aromatic amino acids (260--300 nm). The affinity of photooxidized holoenzyme for glutarate and alpha-methyl aspartate is greatly decreased. On the other hand, photooxidized enzyme retains its ability to bind alpha-alanine and to catalize the transamination half-reaction between alpha-alanine and the bound coenzyme. These findings imply that photooxidation disturbs the binding of the distal carboxyl group of dicarboxylic substrates. This may be due to a localized conformational change induced by destruction of a photoreactive histidine residue at the active site. A role of the histidine residue in transamination reaction is discussed.
D000409:Alanine; D000818:Animals; D001219:Aspartate Aminotransferases / Q000528:radiation effects*; D055598:Chemical Phenomena; D002621:Chemistry; D002645:Chickens / Q000378:metabolism*; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008027:Light; D009206:Myocardium / Q000201:enzymology*; D010084:Oxidation-Reduction; D011732:Pyridoxal Phosphate; D013057:Spectrum Analysis
D011732:Pyridoxal Phosphate; D001219:Aspartate Aminotransferases; D000409:Alanine
Fotookislenie aspartat-transaminazy iz tsitozolia serdtsa kur.