title: Role and location of "protease I" from Escherichia coli.
abstract: Pacaud and Uriel described an enzyme from Escherichia coli ("protease I") that hydrolyzes acetyl phenylalanine naphthyl ester (APNE). We examined the possible involvement of this enzyme in intracellular protein degradation, its subcellular distribution, and its proteolytic activity. Although the APNE-hydrolyzing activity is localized primarily in the periplasm, proteolytic activity against casein was found in the periplasm, membrane, and cytoplasm with similar specific activities. The APNE-hydrolyzing enzyme did not appear to contribute to the proteolytic activity of the periplasm. A mutant deficient in APNE-hydrolyzing activity lacked all activity in the periplasm but showed a slight percentage of residual activity in the cytoplasm. Extracts of such cells were normal in their ability to hydrolyze casein. The mutant was indistinguishable from wild-type cells in its rate of protein degradation during growth or glucose starvation and in the ability to rapidly degrade puromycin-containing polypeptides. Nitrogen starvation, which increased protein breakdown severalfold, affected neither the total amount nor the distribution of APNE-hydrolyzing activity. The mutant showed no defect in its ability to cleave small phenylalanine-containing peptides released during protein degradation. The mutant and wild-type cells are equally able to hydrolyze exogenously supplied phenylalanyl peptides. These experiments suggest that the APNE-hydrolyzing enzyme is not required for protein degradation and that "protease I" is probably not a protease.
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D002918:Chymotrypsin / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology*; D009584:Nitrogen / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D002364:Caseins; D010649:Phenylalanine; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D009584:Nitrogen