Site-directed mutagenesis of the HtrA (DegP) serine protease, whose proteolytic activity is indispensable for Escherichia coli survival at elevated temperatures.
The HtrA(DegP) 48-kDa serine protease of Escherichia coli is indispensable for bacterial survival at elevated temperatures. It contains the amino-acid sequence Gly208AnsSerGlyGlyAlaLeu, which is similar to the consensus sequence GlyAspSerGlyGlyProLys surrounding the active Ser residue of trypsin-like proteases. Mutational alteration of Ser210 eliminated proteolytic activity of HtrA. An identical effect was observed when His105 was mutated. The mutated HtrA were unable to suppress thermosensitivity of the htrA bacteria. These results suggest that Ser210 and His105 may be important elements of the catalytic domain and indicate that the proteolytic activity of HtrA is essential for the survival of cells at elevated temperatures.
Skórko-Glonek
J
J
Department of Biochemistry, University of Gdańsk, Poland.
Wawrzynów
A
A
Krzewski
K
K
Kurpierz
K
K
Lipińska
B
B
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016384:Consensus Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D033903:Periplasmic Proteins; D010957:Plasmids; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid; D012694:Serine; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014357:Trypsin / Q000737:chemistry
D001426:Bacterial Proteins; D006360:Heat-Shock Proteins; D033903:Periplasmic Proteins; D012694:Serine; C061682:DegP protease; D012697:Serine Endopeptidases; D014357:Trypsin