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Literature summary for 6.1.1.20 extracted from

  • Levin, I.; Kessler, N.; Moor, N.; Klipcan, L.; Koc, E.; Templeton, P.; Spremulli, L.; Safro, M.
    Purification, crystallization and preliminary X-ray characterization of a human mitochondrial phenylalanyl-tRNA synthetase (2007), Acta Crystallogr. Sect. F, 63, 761-764.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
mature mitPheRS in Escherichia coli without His-tag, which causes oligomerization of the recombinant enzyme during purification Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant mitPheRS in complex with L-phenylalanine and ATP, hanging or sitting drop vapour diffusion method, 0.001-0.003 ml of protein solution containing 6 mg/ml protein in 20 mM Tris-HCl pH 8.0, 100 mM NaCl, 7 mM MgCl2, 5 mM 2-mercaptoethanol, 1 mM EDTA and 0.0065% NaN3, is mixed with an equal volume of reservoir solution containing 25 mM Tris-HCl pH 8.5, 10 mM MgCl2, 5 mM ATP, 2 mM L-phenylalanine, equilibration against reservoir solution, at 4°C or 19°C, X-ray diffraction structure determination and analysis at 2.2 A resolution Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
Homo sapiens 5739
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
48000
-
recombinant enzyme, gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-phenylalanine + tRNAPhe Homo sapiens
-
AMP + diphosphate + L-phenylalanyl-tRNAPhe
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
mature mitPheRS in Escherichia coli by streptomycin precipitation, ultrafiltration, anion exchange chromatography and heparin affinity chromatography, followed by ammonium sulfate fractionation, hydrophobic interaction chromatography, and gel filtration, to homogeneity Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-phenylalanine + tRNAPhe
-
Homo sapiens AMP + diphosphate + L-phenylalanyl-tRNAPhe
-
?
ATP + L-phenylalanine + tRNAPhe charging of cognate amino acid Homo sapiens AMP + diphosphate + L-phenylalanyl-tRNAPhe
-
?

Subunits

Subunits Comment Organism
monomer 1 * 48000, recombinant enzyme, SDS-PAGE Homo sapiens
More human mitPheRS is a chimera of the bacterial beta-subunit of PheRS and the B8 domain of its beta-subunit, together, the beta-subunit and the RNP-domain, i.e. B8 domain, at the C-terminus form the minimal structural set to construct an enzyme with phenylalanylation activity, overview Homo sapiens

Synonyms

Synonyms Comment Organism
mitochondrial phenylalanyl-tRNA synthetase
-
Homo sapiens
mitPheRS
-
Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
ATP
-
Homo sapiens

pI Value

Organism Comment pI Value Maximum pI Value
Homo sapiens sequence calculation
-
6.4