[In vitro coupling of ATP hydrolysis to proteolysis of ATP site mutant forms of Lon-proteinase from E.coli].
In order to identify amino acid residues involved in ATP hydrolysis by Escherichia coli protease Lon or participating in the signal transduction from the ATPase domain to the proteolytic one, potentially important residues of the ATPase domain were substituted using site-directed mutagenesis, and the properties of the resulting mutant enzymes were studied. It was found that residues K362, T363 (Walker's motif A), and D423 (motif B) are involved in the catalysis of ATP hydrolysis. K362 and T363 also participate in the system of domain-domain coupling, whereas D423 does not play a significant role in this process. Residue D387 is important for ATPase activity; however, it is not a catalytically active residue, as was earlier postulated in the literature. Residue Y493 is also involved in the signal transduction from the ATPase domain to the proteolytic one.
Mel'nikov
E E
EE
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Tsirul'nikov
K B
KB
Ginodman
L M
LM
Rotanova
T V
TV
D049069:ATP-Dependent Proteases; D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000235:genetics; D000255:Adenosine Triphosphate / Q000737:chemistry*; D000494:Allosteric Regulation; D019943:Amino Acid Substitution; D000596:Amino Acids / Q000032:analysis; D002384:Catalysis; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics; D006868:Hydrolysis; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D049070:Protease La; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D013379:Substrate Specificity
D000596:Amino Acids; D017931:DNA Primers; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D000255:Adenosine Triphosphate; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases; C483567:Lon protein, E coli; D049070:Protease La; D000251:Adenosine Triphosphatases
Sopriazhenie gidroliza ATP i proteoliza pri funktsionirovanii in vitro mutagennykh po ATP-asnomy tsentru form Lon-proteinazy E.coli.