title: Allosteric modifier and substrate binding of donkey deoxycytidylate aminohydrolase (EC 3.5.4.12)
abstract: The hexameric allosteric enzyme deoxycytidylate aminohydrolase from donkey spleen is shown by equilibrium dialysis to bind specifically the allosteric inhibitor, dTTP, the activator dCTP, and the substrate analog dAMP each at six sites (the dTTP and dCTP sites may or may not be identical). These conclusions contrast with earlier ones that there were four sites for each effector; reasons for the discrepancy are discussed. With the knowledge of site numbers and the kinetic information from the accompanying paper it is concluded that the kinetic cooperativity of the enzyme excludes a concerted conformational transition mechanism. Amino acid analysis gives a molecular weight of 18,842 Da per subunit, i.e., 113,052 for the hexamer. A new simplified purification of homogeneous enzyme from donkey spleen probably useful for dCMP aminohydrolase from other sources is described.
authors.affiliation: C.N.R., Institute of Protein Biochemistry and Enzymology, Napoli, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Nucci
authors.forename: C A
authors.initials: CA
authors.lastname: Raia
authors.forename: C
authors.initials: C
authors.lastname: Vaccaro
authors.forename: M
authors.initials: M
authors.lastname: Rossi
authors.forename: E P
authors.initials: EP
authors.lastname: Whitehead
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives* / Q000378:metabolism; D000494:Allosteric Regulation; D000495:Allosteric Site; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003844:DCMP Deaminase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D003845:Deoxycytosine Nucleotides / Q000378:metabolism*; D004789:Enzyme Activation; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D010527:Perissodactyla; D011487:Protein Conformation; D013154:Spleen / Q000201:enzymology; D013942:Thymine Nucleotides / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D003845:Deoxycytosine Nucleotides; D046911:Macromolecular Substances; D013942:Thymine Nucleotides; C024107:2'-deoxycytidine 5'-triphosphate; C016358:2,4-diamino-5-adamantyl-6-methylpyrimidine; D003844:DCMP Deaminase; D000218:Adamantane; C024157:thymidine 5'-triphosphate