Abstract
1. The time-course of tryptic hydrolysis of aspartate transcarbamoylase (aspartate carbamoyltransferase, EC 2.1.3.2) was followed by activity measurements in the presence and absence of allosteric effectors, and by polyacrylamide-gel electrophoresis. 2. Two proteins with enzyme activity are formed in this way from native enzyme, and the isolation and some properties of these species are reported. The larger protein (10.6S) resembles native enzyme in that it contains regulatory subunits and is sensitive to allosteric effectors, as well as in a more detailed kinetic investigation. It appears from the time-course of tryptic digestion to be an intermediate in the formation of a catalytic subunit (5.5S) which is similar to, but not identical with, the catalytic subunit produced by mercurial treatment of the native enzyme. 3. Sodium dodecyl sulphate–polyacrylamide-gel electrophoresis of the different enzyme forms demonstrates that trypsin can hydrolyse bonds in the catalytic polypeptide chains as well as completely remove the regulatory polypeptide chains. 4. Both preparations of catalytic subunit can recombine with regulatory subunit to form enzymes which resemble the native enzyme in being activated by ATP, although they do not appear to be inhibited by CTP. 5. This study is consistent with the models of the enzyme that propose that the catalytic subunits are held together in the native enzyme by three pairs of regulatory polypeptide chains.
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