Basis for the observed fluctuation of carboxypeptidase II activity during the cell cycle in BUG 6, a temperature-sensitive division mutant of Escherichia coli.

Diaminopimelyl-d-alanyl carboxypeptidase (carboxypeptidase II) is most active at the time of division, whether measured in toluene-treated cells of Escherichia coli K-12 strain D11-1, fractionated by size, or in toluene-treated cells of the temperature-sensitive division mutant, BUG 6 (B. D. Beck and J. T. Park, 1976). The present investigation has now shown that, under conditions that permit division, the increased carboxypeptidase II activity in toluenetreated cells of BUG 6 is probably not due to protein synthesis. Although dividing cells are more permeable than nondividing cells, permeability differences are not sufficient to account for the changes in carboxypeptidase II activity. Thus, in the toluene-treated nondividing cells, carboxypeptidase II is present, but its activity is masked, which suggests the presence of an inhibitor. Another striking difference between nondividing and dividing cells is that carboxypeptidase II is much more readily released from dividing cells by both tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetic acid and toluene treatment. Carboxypeptidase II was partially purified and found to be an 86,000-molecular-weight protein consisting of two 43,000-molecular-weight polypeptides. Tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetic acid treatment of nondividing cells releases less than 10% of the carboxypeptidase II and other periplasmic proteins that are releasable from dividing cells.