Literature summary for 3.4.21.53 extracted from

An, Y.J.; Na, J.H.; Kim, M.I.; Cha, S.S.
Structural basis for the ATP-independent proteolytic activity of LonB proteases and reclassification of their AAA+ modules (2015), J. Microbiol., 53, 711-717 .

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Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His-tagged enzyme with deletion of a membrane-anchoring region (residues 134-170) in Escherichia coli strain BL21(DE3)	Thermococcus onnurineus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified isolated AAA+ module of LonB protease, mixing of 0.001 ml of 12 mg/ml protein solution in 10 mM Tris-HCl, pH 7.5, 100 mM NaCl, and 1 mM DTT, with 0.001 ml of reservoir solution containing 0.2 M potassium chloride, 0.01 M magnesium acetate, 0.05 M tri-sodium citrate dihydrate, pH 4.5, 12% PEG 4000, and 5% w/v n-dodecyl-beta-D-maltoside, 22°C, X-ray diffraction structure determination and analysis at 2.03 A resolution	Thermococcus onnurineus

Crystallization (Comment)

Organism

purified isolated AAA+ module of LonB protease, mixing of 0.001 ml of 12 mg/ml protein solution in 10 mM Tris-HCl, pH 7.5, 100 mM NaCl, and 1 mM DTT, with 0.001 ml of reservoir solution containing 0.2 M potassium chloride, 0.01 M magnesium acetate, 0.05 M tri-sodium citrate dihydrate, pH 4.5, 12% PEG 4000, and 5% w/v n-dodecyl-beta-D-maltoside, 22°C, X-ray diffraction structure determination and analysis at 2.03 A resolution

Thermococcus onnurineus

Organism

Organism	UniProt	Comment	Textmining
Thermococcus onnurineus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant truncated His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, the enzyme is reduced to the AAA+ domain by proteolytic treatment with trypsin followed by gel filtration	Thermococcus onnurineus

Subunits

Subunits	Comment	Organism
hexamer	-	Thermococcus onnurineus

Synonyms

Synonyms	Comment	Organism
lonB protease	-	Thermococcus onnurineus
TonLonB	-	Thermococcus onnurineus

Cofactor

Cofactor	Comment	Organism	Structure
ATP	activates, the ATP-binding site is situated at the interface between the alpha/beta-subdomain and the alpha-subdomain. Motifs for ATP hydrolysis and nucleotide sensing are well conserved at the interface. Structure, detailed overview	Thermococcus onnurineus

General Information

General Information	Comment	Organism
evolution	structural comparison of AAA+ modules between LonA and LonB reveals that the AAA+ modules of Lon proteases are separated into two distinct clades depending on their structural features. The H2-insert clade is characterized by the presence of both the PS-1 insertion and the H2 insertion. In the H2-insert clade, no member has an additional insertion, like Ins1, between alpha4 and beta2 in the RFC-B AAA+ module. Therefore, it is not appropriate to classify the AAA+ module of TonLonB as the H2-insert clade. Alternatively, the AAA+ module of LonB belongs to a new clade, named H2 & Ins1 insert clade is proposed. The AAA+ module of LonB belongs to the H2 & Ins1 insert clade (HINS clade), while the AAA+ module of LonA is a member of the HCLR clade	Thermococcus onnurineus
additional information	crystal structure determination of AAA+ module. Structural basis for the ATP-independent proteolytic activity of LonB proteases and reclassification of their AAA+ modules, overview. The isolated AAA+ module, having no bound nucleotide, adopts a conformation virtually identical to the ADP-bound conformation of AAA+ modules in the hexameric structure of TonLonB. Despite the conservation of functional motifs, the iAAA+ module has no ATPase activity. In the hexameric conformation, an arginine finger (Arg311) in one AAA+ module stabilizes negative charge of gamma-phosphate of ATP bound to the adjacent AAA+ module, which is essential for ATP hydrolysis	Thermococcus onnurineus