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Literature summary extracted from
- Zinc is the molecular switch that controls the catalytic cycle of bacterial leucyl-tRNA synthetase (2015), J. Inorg. Biochem., 142, 59-67 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 6.1.1.4 | gene leuS, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 from plasmid pRWecLeuRS | Escherichia coli |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 6.1.1.4 | C159A | site-directed mutagenesis, structure comparison with the wild-type | Escherichia coli |
| 6.1.1.4 | C176A | site-directed mutagenesis, structure comparison with the wild-type | Escherichia coli |
| 6.1.1.4 | C179A | site-directed mutagenesis, structure comparison with the wild-type | Escherichia coli |
| 6.1.1.4 | additional information | C159A, C176A and C179A disassociate from Zn2+ much more readily than wild-type LeuRS, the wild-type LeuRS binds more tightly to Zn2+ than do C159A, C176A or C179A | Escherichia coli |
| 6.1.1.4 | T252Y | site-directed mutagenesis, an editing defective mutant | Escherichia coli |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 6.1.1.4 | additional information | - |
additional information | determination of dissociation constants of Zn2+ from LeuRS enzymes | Escherichia coli |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 6.1.1.4 | Mg2+ | required | Escherichia coli |  |
| 6.1.1.4 | Zn2+ | required, zinc is the molecular switch that controls the catalytic cycle of bacterial leucyl-tRNA synthetase. A specialized zinc-binding domain 1 (ZN-1), when associated with Zn2+, assumes a rigid architecture that is stabilized by thiol groups from the residues C159, C176, and C179. When LeuRS is in the aminoacylation complex, these cysteine residues forman equilateral planar triangular configuration with Zn2+, but when LeuRS transitions to the editing conformation, this geometric configuration breaks down. The wild-type enzyme binds Zn2+ to a greater extent than any of the mutant LeuRSs | Escherichia coli | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 6.1.1.4 | ATP + L-leucine + tRNALeu | Escherichia coli | - |
AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 6.1.1.4 | Escherichia coli | P07813 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 6.1.1.4 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 by nickel affinity chromatography | Escherichia coli |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 6.1.1.4 | ATP + L-leucine + tRNALeu | - |
Escherichia coli | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 6.1.1.4 | More | structure comparison of the Zn-1 domain in editing and in aminoacylation conformations, overview | Escherichia coli |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 6.1.1.4 | b0642 | - |
Escherichia coli |
| 6.1.1.4 | JW0637 | - |
Escherichia coli |
| 6.1.1.4 | Leucyl-tRNA synthetase | - |
Escherichia coli |
| 6.1.1.4 | LeuRS | - |
Escherichia coli |
| 6.1.1.4 | leuS | - |
Escherichia coli |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 6.1.1.4 | 30 | - |
assay at | Escherichia coli |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 6.1.1.4 | 7.5 | - |
assay at | Escherichia coli |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 6.1.1.4 | ATP | - |
Escherichia coli | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 6.1.1.4 | evolution | enzyme leucyl-tRNA synthetase is part of the aminoacyl-tRNA synthetase (aaRS) family | Escherichia coli |
| 6.1.1.4 | additional information | there are two catalytic sites, the leucylation site housed within the aminoacylation domain and the hydrolytic deacylation site housed within the CP1 editing domain, the ZN-1 domain is known to play an essential structural role in stabilizing the Leu-Amp adenylate. Structural analysis of LeuRS enzymes using Fourier transform infrared spectroscopy (FTIR), and homology modeling of LeuRS in the editing conformation, visualizing the ZN-1 domain, by using the structure in editing conformation of the LeuRS enzyme from Thermus thermophilus, PDB ID 1OBH as template, overview | Escherichia coli |
| 6.1.1.4 | physiological function | Escherichia coli leucyl-tRNA synthetase (LeuRS) is an essential multi-domain metalloenzyme that aminoacylates tRNALeu with leucine. Enzyme LeuRS is an essential enzyme that relies on specialized domains to facilitate the aminoacylation reaction. Structural changes within the ZN-1 domain play a central role in LeuRS's catalytic cycle. The enzyme performs a Zn2+ dependent translocation mechanism for charged tRNALeu, Zn2+ is an architectural cornerstone of the ZN-1 domain and that without its geometric coordination the domain collapses. Residues C159, C176 and C179 coordinate Zn2+ and that this interaction is essential for leucylation to occur, but is not essential for deacylation | Escherichia coli |
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