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Literature summary for 3.4.21.53 extracted from

  • Kudzhaev, A.; Andrianova, A.; Serova, O.; Arkhipova, V.; Dubovtseva, E.; Rotanova, T.
    The effect of mutations in the inserted domain of ATP-dependent Lon protease from E. coli on the enzyme function (2015), Russ. J. Bioorg. Chem., 41, 518-524 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene lon, recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
additional information analysis of the propensity of C-His-Lon and mutant enzymes Lon-R164A, Lon-R192A, and Lon-Y294A for autodegradation reveals that Lon-Y294A is most prone to autolysis. Slight autolysis of the intact enzyme and the mutant forms of Lon-R164A and Lon-R192A is observed only in the absence of nucleotides Escherichia coli
R164A site-directed mutagenesis, mutation of a HI(CC) domain residue, helix H3, the mutant shows highly reduced ATPase activity in presence of beta-casein compared to the wild-type Escherichia coli
R192A site-directed mutagenesis, mutation of a HI(CC) domain residue, the mutant shows highly reduced ATPase activity in presence of beta-casein compared to the wild-type Escherichia coli
Y294A site-directed mutagenesis, mutation of a HI(CC) domain residue, the mutant shows highly reduced ATPase activity in presence of beta-casein compared to the wild-type Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
ADP free ADP inhibits all forms of Ec-Lon, while a complex of ADP with magnesium ions (ADP-Mg) activates C-His-Lon and Lon-R164A and inhibits the mutant forms Lon R192A and Lon-Y294A Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required, activates Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A9M0
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-casein + H2O
-
Escherichia coli ?
-
?

Subunits

Subunits Comment Organism
homohexamer
-
Escherichia coli
More enzyme Ec-Lon subunit includes an ATPase component and a proteolytic component (AAA+ module and P-domain, respectively), as well as a noncatalytic region formed by the N-terminal (N) domain and an inserted alpha-helical (HI(CC)) domain. This region is unique for AAA+ proteins. The C-terminal part of the HI(CC) domain has an allosteric effect on the efficiency of the functioning of both ATPase and proteolytic sites of the enzyme, while the coiled-coil (CC) fragment of this domain interacts with the protein substrate. The HI(CC) domain is not essential for the formation of the ATPase center of Ec-Lon protease, but still has an effect on the functional efficiency of this center. Detailed analysis and comparisons of primary and secondary structures of the HI(CC) domain in AAA* proteases, overview Escherichia coli

Synonyms

Synonyms Comment Organism
ATP-dependent protease LonA
-
Escherichia coli
Ec-Lon
-
Escherichia coli
lonA
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
-
Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.1
-
-
Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP dependent on Escherichia coli

General Information

General Information Comment Organism
evolution LonA from Escherichia coli belongs to the superfamily of AAA+ proteins, family S16 of proteases. The presence of an extended variable N-terminal region preceding the AAA+ modules is a characteristic feature of LonA proteases distinguishing them from other AAA+ proteins Escherichia coli
additional information the C-terminal part of the HI(CC) domain has an allosteric effect on the efficiency of the functioning of both ATPase and proteolytic sites of the enzyme, while the coiled-coil (CC) fragment of this domain interacts with the protein substrate. Analysis of the propensity of C-His-Lon and mutant enzymes Lon-R164A, Lon-R192A, and Lon-Y294A for utodegradation reveals that Lon-Y294A is most prone to autolysis. Slight autolysis of the intact enzyme and the mutant forms of Lon-R164A and Lon-R192A is observed only in the absence of nucleotides Escherichia coli
physiological function LonA from Escherichia coli plays a key role in the quality control system of the cell proteome. It destroys abnormal and defective polypeptides, as well as a number of regulatory proteins, according to a processive degradation mechanism Escherichia coli