TY - JOUR
T1 - Mutations in the proteolytic domain of Escherichia coli protease Lon impair the ATPase activity of the enzyme
AU - Starkova, Natalie N.
AU - Koroleva, Ekaterina P.
AU - Rumsh, Lev D.
AU - Ginodman, Lev M.
AU - Rotanova, Tatyana V.
N1 - Funding Information:
We are grateful to Dr. N.S. Bystrov for the synthesis of oligonucleotides. This work was supported in part by the Russian Foundation for Basic Research (project no. 93-04-6201) and the International Science Foundation (project NF8000).
PY - 1998/1/30
Y1 - 1998/1/30
N2 - Conserved residues of the proteolytic domain of Escherichia coli protease Lon, putative members of the classic catalytic triad (H665, H667, D676, and D743) mere identified by comparison of amino acid sequences of Lon proteases. Mutant enzymes containing substitutions D676N, D743N, H665Y, and H667Y were obtained by site-directed mutagenesis. The mutant D743N retained the adenosine triphosphate (ATP)-dependent proteolytic activity, thereby indicating that D743 does not belong to the catalytic site. Simultaneously, the mutants D676N, H665Y, add H667Y lost the capacity for hydrolysis of protein substrates. The ATPase activity of these three mutants was decreased by more than an order of magnitude, which suggests a close spatial location of the ATPase and proteolytic active sites and their tight interaction in the process of protein degradation.
AB - Conserved residues of the proteolytic domain of Escherichia coli protease Lon, putative members of the classic catalytic triad (H665, H667, D676, and D743) mere identified by comparison of amino acid sequences of Lon proteases. Mutant enzymes containing substitutions D676N, D743N, H665Y, and H667Y were obtained by site-directed mutagenesis. The mutant D743N retained the adenosine triphosphate (ATP)-dependent proteolytic activity, thereby indicating that D743 does not belong to the catalytic site. Simultaneously, the mutants D676N, H665Y, add H667Y lost the capacity for hydrolysis of protein substrates. The ATPase activity of these three mutants was decreased by more than an order of magnitude, which suggests a close spatial location of the ATPase and proteolytic active sites and their tight interaction in the process of protein degradation.
KW - Active site
KW - Adenosine triphosphate-dependent proteolysis
KW - Escherichia coli
KW - Protease Lon
KW - Site-directed mutagenesis
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U2 - 10.1016/S0014-5793(98)00012-X
DO - 10.1016/S0014-5793(98)00012-X
M3 - Article
C2 - 9490010
AN - SCOPUS:0032579436
SN - 0014-5793
VL - 422
SP - 218
EP - 220
JO - FEBS Letters
JF - FEBS Letters
IS - 2
ER -