title: Characterization of a beta-galactosidase formed between a complementary protein and a peptide synthesized de novo.
abstract: In a cell-free system, phi80dlac can be transcribed, and the resulting ribonucleic acid can be translated to yield a product which interacts with an enzymatically inactive z protein to produce active enzyme. The inactive z protein is produced by Escherichia coli strain 21, which contains a deletion in the first part of the gene for beta-galactosidase and appears to exist as a dimer. The enzyme formed in the cell-free system appears to be composed of one strain 21 z protein dimer and one newly synthesized polypeptide chain with a molecular weight of about 3 x 10(4). The estimated size of this complementing segment is in good agreement with Ullmann, Jacob, and Monod's estimate of the size of the alpha region of beta-galactosidase. Using alpha fragments produced by autoclaving or guanidine treatments, we found that the active portion of alpha seems to be smaller than the full alpha region. We also found, using alpha produced by the autoclaving technique, that active dimer undergoes conversion to tetramer as the amount of alpha is increased. Evidently, the binding of alpha favors this conversion, but it is unlikely that the conversion of dimer to tetramer per se results in increased enzyme activity.
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000378:metabolism; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004269:DNA, Bacterial / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology*; D005696:Galactosidases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D007930:Leucine / Q000378:metabolism; D008967:Molecular Biology; D008970:Molecular Weight; D009154:Mutation; D010452:Peptide Biosynthesis; D013698:Templates, Genetic; D014316:Tritium
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D014316:Tritium; D005696:Galactosidases; D007930:Leucine