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Literature summary extracted from

  • Oka, M.; Takegawa, K.; Kimura, Y.
    Enzymatic characterization of a class II lysyl-tRNA synthetase, LysS, from Myxococcus xanthus (2015), Arch. Biochem. Biophys., 579, 33-39 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.6 gene lysS, recombinant expression of His-tagged enzyme in Escherichia coli Myxococcus xanthus

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.1.1.6 ADP the addition of ADP inhibits the diadenosine tetraphosphate synthesis by Ap4A synthase activity of LysS. Ap4A production decreases by 22% and 52% at 1 mM and 5 mM ADP, respectively. In the presence of ADP, Ap3A is not synthesized by LysS during 30 min of incubation. ADP may act as a competitive inhibitor of LysS in the first step reaction, i.e. lysyl-AMP formation from lysine and ATP Myxococcus xanthus
6.1.1.6 Zn2+ inhibits enzyme activity in the presence of Mn2+ Myxococcus xanthus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.1.1.6 Mg2+ required Myxococcus xanthus
6.1.1.6 Mn2+ the enzyme requires Mn2+ for the synthesis of diadenosine tetraphosphate (Ap4A) Myxococcus xanthus
6.1.1.6 additional information Zn2+ is not required for diadenosine tetraphosphate (Ap4A) synthesis by LysS Myxococcus xanthus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.1.1.6 68000
-
recombinant His-tagged enzyme, gel filtration Myxococcus xanthus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.1.1.6 ATP + L-lysine + tRNALys Myxococcus xanthus
-
AMP + diphosphate + L-lysyl-tRNALys
-
?
6.1.1.6 ATP + L-lysine + tRNALys Myxococcus xanthus DK 1622
-
AMP + diphosphate + L-lysyl-tRNALys
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.6 Myxococcus xanthus Q1D380
-
-
6.1.1.6 Myxococcus xanthus DK 1622 Q1D380
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.6 recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, dialysis, EDTA treatment, and again dialysis Myxococcus xanthus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.6 ATP + L-lysine + tRNALys
-
Myxococcus xanthus AMP + diphosphate + L-lysyl-tRNALys
-
?
6.1.1.6 ATP + L-lysine + tRNALys
-
Myxococcus xanthus DK 1622 AMP + diphosphate + L-lysyl-tRNALys
-
?
6.1.1.6 additional information lysyl-tRNA synthetases efficiently produce diadenosine tetraphosphate (Ap4A) from lysyl-AMP with ATP in the absence of tRNA. When incubated with ATP, Mn2+, lysine, and inorganic diphosphatase, LysS first produces Ap4A and ADP, then converts Ap4A to diadenosine triphosphate (Ap3A), and finally converts Ap3A to ADP, the end product of the reaction. Recombinant LysS retains Ap4A synthase activity without lysine addition. When incubated with Ap4A (minus pyrophosphatase), LysS converts Ap4A mainly ATP and AMP, or ADP in the presence or absence of lysine, respectively. LysS converts Ap4A to yield ATP and AMP when Mn2+ and lysine are present, formation of ATP is higher than that of AMP. LysS hydrolyzes Ap4A symmetrically yielding ADP Myxococcus xanthus ?
-
?
6.1.1.6 additional information lysyl-tRNA synthetases efficiently produce diadenosine tetraphosphate (Ap4A) from lysyl-AMP with ATP in the absence of tRNA. When incubated with ATP, Mn2+, lysine, and inorganic diphosphatase, LysS first produces Ap4A and ADP, then converts Ap4A to diadenosine triphosphate (Ap3A), and finally converts Ap3A to ADP, the end product of the reaction. Recombinant LysS retains Ap4A synthase activity without lysine addition. When incubated with Ap4A (minus pyrophosphatase), LysS converts Ap4A mainly ATP and AMP, or ADP in the presence or absence of lysine, respectively. LysS converts Ap4A to yield ATP and AMP when Mn2+ and lysine are present, formation of ATP is higher than that of AMP. LysS hydrolyzes Ap4A symmetrically yielding ADP Myxococcus xanthus DK 1622 ?
-
?

Subunits

EC Number Subunits Comment Organism
6.1.1.6 monomer 1 x 58000, about, recombinant His-tagged enzyme, sequence calculation, 1 * 60000, recombinant His-tagged enzyme, SDS-PAGE Myxococcus xanthus

Synonyms

EC Number Synonyms Comment Organism
6.1.1.6 Ap4A synthase
-
Myxococcus xanthus
6.1.1.6 class II lysyl-tRNA synthetase
-
Myxococcus xanthus
6.1.1.6 lysS
-
Myxococcus xanthus
6.1.1.6 MXAN_4731
-
Myxococcus xanthus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.1.1.6 50
-
-
Myxococcus xanthus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
6.1.1.6 30 60 the relative activity of LysS at 30-60°C is 49%, 77%, or 45%, respectively, compared to the activity at the optimal temperature of 50°C Myxococcus xanthus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.1.1.6 8 9
-
Myxococcus xanthus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
6.1.1.6 7 9 optimal activity at pH 8.0-9.0, 80% of maximal activity at pH 7.0 Myxococcus xanthus

Cofactor

EC Number Cofactor Comment Organism Structure
6.1.1.6 ATP
-
Myxococcus xanthus

General Information

EC Number General Information Comment Organism
6.1.1.6 evolution the lysyl-tRNA synthetase of Mycobacterium smegmatis belongs to the class II amino acid tRNA synthetases Myxococcus xanthus
6.1.1.6 physiological function activity of most class II aminoacyl-tRNA synthetases absolutely requires the presence of added cognate amino acids Myxococcus xanthus