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Literature summary for 6.1.1.7 extracted from

  • Banerjee, B.; Banerjee, R.
    Urea unfolding study of E. coli alanyl-tRNA synthetase and its monomeric variants proves the role of C-terminal domain in stability (2015), J. Amino Acids, 2015, 805681 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.1.1.7

Cloned(Commentary)

Cloned (Comment) Organism
gene alaS, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
G674D site-directed mutagenesis, a point mutation in the C-terminal domain, the mutation produces a monomeric variant with a fivefold reduced aminoacylation activity compared to the wild-type enzyme Escherichia coli
additional information a truncated enzyme comparising the N-terminal 700 amino acids forms a monomeric protein possessing similar aminoacylation activity like the wild-type enzyme, and construction of N461 truncated enzyme. Aminoacylation activity assay after refolding from denatured state reveals that the monomeric mutants are unable to regain their activity, whereas the dimeric full-length alaRS gets back similar activity as the native enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information thermodynamics of wild-type and mutant enzymes Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-alanine + tRNAAla Escherichia coli
-
 AMP + diphosphate + L-alanyl-tRNAAla
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P00957
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzmyes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Escherichia coli

Renatured (Commentary)

Renatured (Comment) Organism
after refolding from denatured state, the dimeric full-length alaRS gets back similar activity as the native enzyme, while the monomeric, truncated or point mutants do not, secondary structure analysis, unfolding profiles, overview Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-alanine + tRNAAla
-
 Escherichia coli AMP + diphosphate + L-alanyl-tRNAAla
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 96000, analytical ultracentrifugation Escherichia coli
More alanyl-tRNA synthetase exists as a dimer in its native form and the C-terminal coiled-coil part plays an important role in the dimerization process. Dimerization is one of the key regulatory factors that is important in the proper folding and stability of Escherichia coli alaRS Escherichia coli

Synonyms

Synonyms Comment Organism
Alanyl-tRNA synthetase
-
 Escherichia coli
AlaRS
-
 Escherichia coli

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Escherichia coli

General Information

General Information Comment Organism
evolution the enzyme belongs to the class IIa aminoacyl-tRNA synthestase family Escherichia coli