Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | magnesium-dependent activation and hexamerization of the Lon AAA+ protease | Meiothermus taiwanensis |
Cloned (Comment) | Organism |
---|---|
gene lonA1, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Meiothermus taiwanensis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant full-length wild-type enzyme LonA in complex with bortezomib and Mg2+, and in apoform with Mg2+, hexameric and dodecameric enzyme crystals, AAAP-Mg2+-bortezomib complex by hanging drop vapor diffusion method, for the apoenzyme-Mg2+ crystals, 400 nl of 8 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 100 mM NaCl, and 10 mM MgCl2, is mixed with 400 nl of well solution containing 20% PEG 3000, 0.1 M HEPES, pH 7.5, and 0.2 M NaCl, for the complex crystals, mixing of 0.001 ml of protein-inhibitor solution with 0.001 ml of well solution containing 0.2 M sodium citrate, pH 6.5, and 10% PEG 3350, 22°C, X-ray diffraction structure determination and analysis at 1.85 A and 3.05 A resolution, respectively, molecular replacement | Meiothermus taiwanensis |
Protein Variants | Comment | Organism |
---|---|---|
E423Q | site-directed mutagenesis, structure determination | Meiothermus taiwanensis |
I398G | site-directed mutagenesis | Meiothermus taiwanensis |
additional information | generation of truncated enzyme versions, AAAP(residues 295-793) and AP(residues 492-793) | Meiothermus taiwanensis |
S678A | site-directed mutagenesis | Meiothermus taiwanensis |
Y397G | site-directed mutagenesis | Meiothermus taiwanensis |
Y397G/I398G | site-directed mutagenesis, the mutant fails to degrade alpha-casein with Mg2+ and ATP. The Mg2+-activated double mutant showed wild-type-like ATP-independent proteolysis | Meiothermus taiwanensis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
bortezomib | enzyme-bound structure analysis | Meiothermus taiwanensis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | magnesium-dependent activation and hexamerization of the Lon AAA+ protease. Mg2+-binding site structure analysis, Mg2+ is bound in the proteolytic groove | Meiothermus taiwanensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Meiothermus taiwanensis | A0A059VAZ3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Meiothermus taiwanensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-casein + H2O | - |
Meiothermus taiwanensis | ? | - |
? | |
Ig2 + H2O | - |
Meiothermus taiwanensis | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | Mg2+-activated LonA forms an open hexameric chamber without nucleotide | Meiothermus taiwanensis |
More | Mg2+-dependent activation and hexamerization of the Lon AAA+ protease. Role of the protease domains in the oligomerization and activity of LonA, overview | Meiothermus taiwanensis |
Synonyms | Comment | Organism |
---|---|---|
AAAP | - |
Meiothermus taiwanensis |
Lon AAA+ protease | - |
Meiothermus taiwanensis |
lonA | - |
Meiothermus taiwanensis |
MtaLonA | - |
Meiothermus taiwanensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
AMPPNP | adenylylimidodiphosphate, is not as effective as ATP, slight activation | Meiothermus taiwanensis | |
ATP | activates | Meiothermus taiwanensis | |
additional information | Mg2+-activated LonA degrades unfolded protein or peptides without ATP | Meiothermus taiwanensis |
General Information | Comment | Organism |
---|---|---|
additional information | Mg2+-activated LonA can operate as a diffusion-based chambered protease to degrade unstructured protein and peptide substrates efficiently in the absence of ATP. Mg2+-dependent remodeling of a substrate-binding loop and a potential metal-binding site near the Ser-Lys catalytic dyad, supported by biophysical binding assays and molecular dynamics simulations, domain arrangement and structural modeling, overview | Meiothermus taiwanensis |
physiological function | the Lon AAA+ protease (LonA) plays important roles in protein homeostasis and regulation of diverse biological processes. Proposed Mg2+- and nucleotide-dependent assembly pathway of LonA, overview | Meiothermus taiwanensis |