Organization of a multifunctional protein in pyrimidine biosynthesis. A domain hypersensitive to proteolysis.
When the multifunctional protein that catalyses the first three steps of pyrimidine biosynthesis in hamster cells is treated with staphylococcal V8 proteinase, a single cleavage takes place. The activities of carbamoyl-phosphate synthetase (EC 6.3.5.5), aspartate carbamoyltransferase (EC 2.1.3.2) and dihydro-orotase (EC 3.5.2.3) and the allosteric inhibition by UTP are unaffected. One fragment, of Mr 182000, has the first and third enzyme activities, whereas the other fragment, of Mr 42000, has aspartate carbamoyltransferase activity and an aggregation site. A similar small fragment is observed in protein digested with low concentrations of trypsin. A similar large fragment is seen after digestion with trypsin and as the predominating form of this protein in certain mutants defective in pyrimidine biosynthesis. These results indicate that a region located adjacent to the aspartate carbamoyltransferase domain is hypersensitive to proteinase action in vitro and may also be sensitive to proteolysis in vivo.
D000494:Allosteric Regulation; D000818:Animals; D001221:Aspartate Carbamoyltransferase; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing); D002460:Cell Line; D006224:Cricetinae; D004080:Dihydroorotase; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D008647:Mesocricetus; D009097:Multienzyme Complexes / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D010447:Peptide Hydrolases; D011743:Pyrimidines / Q000096:biosynthesis*; D012697:Serine Endopeptidases; D014357:Trypsin
C033626:CAD trifunctional enzyme; D009097:Multienzyme Complexes; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011743:Pyrimidines; D001221:Aspartate Carbamoyltransferase; D010450:Endopeptidases; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin; D004080:Dihydroorotase; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing); C030986:pyrimidine