Cloned (Comment) | Organism |
---|---|
gene lonC, cloning and expression of N-terminally His6-tagged residues 35-387 of the N-terminal domain of the mutant L91M/L188M/I359M enzyme with a TEV protease-cleavage site as selenomethionine-labeled protein in Escherichia coli strain B834(DE3) | Meiothermus taiwanensis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant selenomethionine-labeled N-terminal domain mutant L91M/L188M/I359M , mixing of 0.001 ml of 8 mg/ml protein in 10 mM HEPES, pH 7.5, 100 mM NaCl, 10% v/v glycerol, 1 mM DTT, with 0.001 ml of well solution containing consisting of 400 mM lithium sulfate, 10 mM nickel chloride, 100 mM Tris-HCl, pH 8.4, 22°C, 7 days to 3 months, Selected crystals are further dehydrated for 3 days in sitting drops equilibrated against 0.7-1.2 M lithium sulfate, X-ray diffraction structure determination and analysis at 3.1-3.4 A resolution, modelling | Meiothermus taiwanensis |
Protein Variants | Comment | Organism |
---|---|---|
L91M/L188M/I359M | site-directed mutagenesis, the three mutations are introduced into the wild-type sequence to facilitate de novo phasing | Meiothermus taiwanensis |
additional information | generation of deletion mutations MtaLonCdelta (removing residues 506-511) and MtaLonCDELTAHHE (replacing residues 118-205 with a triglycine linker) by a PCR-based strategy | Meiothermus taiwanensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Meiothermus taiwanensis | C9DRU9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant selenomethionine-labeled N-terminally His6-tagged N-terminal domain of mutant L91M/L188M/I359M enzyme from Escherichia coli strain B834(DE3) by nickel affinity chromatography, tag cleavage by TEV protease, anion exchange chromatography, and gel filtration | Meiothermus taiwanensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Abz-QLRSLNGEWRFAWFPAPEAV[Tyr(3-NO2)]A + H2O | i.e. F-beta20-Q peptide, a synthetic fluorogenic peptide | Meiothermus taiwanensis | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | quaternary enzyme structure, modelling, overview | Meiothermus taiwanensis |
Synonyms | Comment | Organism |
---|---|---|
LonC | - |
Meiothermus taiwanensis |
LonC protease | - |
Meiothermus taiwanensis |
MtaLonC | - |
Meiothermus taiwanensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
assay at | Meiothermus taiwanensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Meiothermus taiwanensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | ATP-dependent protease, the enzyme contains an AAA-like domain that lacks the conserved ATPase motifs. The LonC AAA-like domain is inserted with a large domain predicted to be largely alpha-helical, the unique Lon-insertion domain is disordered in the full-length crystal structure of Meiothermus taiwanensis LonC | Meiothermus taiwanensis |
General Information | Comment | Organism |
---|---|---|
evolution | although Lon is originally identified as an ATP-dependent protease with fused AAA+ (ATPases associated with diverse cellular activities) and protease domains, analyses have recently identified LonC as a class of Lon-like proteases with no intrinsic ATPase activity. In contrast to the canonical ATP-dependent Lon present in eukaryotic organelles and prokaryotes, LonC contains an AAA-like domain that lacks the conserved ATPase motifs | Meiothermus taiwanensis |
additional information | the N-terminal substrate-recognition domain of a LonC protease exhibits structural and functional similarity to cytosolic chaperones | Meiothermus taiwanensis |
physiological function | the Lon-insertion domain of LonC is involved both in Skplike chaperone activity and in recognition of unfolded protein substrates, structure of Lon-insertion domain is remarkably similar to the tentacle-like prong of the periplasmic chaperone Skp | Meiothermus taiwanensis |