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Literature summary extracted from

  • Ramesh, V.; Gite, S.; RajBhandary, U.L.
    Functional interaction of an arginine conserved in the sixteen amino acid insertion module of Escherichia coli methionyl-tRNA formyltransferase with determinants for formylation in the initiator tRNA (1998), Biochemistry, 37, 15925-15932.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.2.9 expression of wild-type and of R42K, R42L and G41R/R42L mutant enzymes in Escherichia coli JM109 as His6-tagged proteins Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.2.9
-
 Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.1.2.9 G41D mutant enzyme with decreased activity Escherichia coli
2.1.2.9 G41K mutant enzyme with decreased activity Escherichia coli
2.1.2.9 G41L mutant enzyme with decreased activity Escherichia coli
2.1.2.9 G41R G41R/R42L double mutant with low activity, G41R: decreased activity Escherichia coli
2.1.2.9 R38K mutant enzyme with 2.2fold increased Km, no effect on kcat Escherichia coli
2.1.2.9 R38L mutant enzyme with up to 12fold lower specific activity, 6fold increased Km and 5fold decreased kcat Escherichia coli
2.1.2.9 R42K mutant enzyme with lower activity, 16fold increased Km Escherichia coli
2.1.2.9 R42L single mutant and G41R/R42L double mutant with low activity, R42L: 17fold increased Km and 79fold decreased kcat Escherichia coli
2.1.2.9 R42Q mutant enzyme with lower activity, 16-17fold increased Km Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.2.9 additional information
-
 additional information values for wild-type and mutant enzyme with wild-type and mutant initiator tRNAs Escherichia coli
2.1.2.9 0.00054
-
 L-methionyl-tRNAfMet wild-type enzyme Escherichia coli
2.1.2.9 0.00066
-
 L-methionyl-tRNAfMet G41R mutant enzyme Escherichia coli
2.1.2.9 0.00102
-
 L-methionyl-tRNAfMet G41K mutant enzyme Escherichia coli
2.1.2.9 0.00121
-
 L-methionyl-tRNAfMet R38K mutant enzyme Escherichia coli
2.1.2.9 0.00162
-
 L-methionyl-tRNAfMet G41L mutant enzyme Escherichia coli
2.1.2.9 0.003
-
 L-methionyl-tRNAfMet G41D mutant enzyme Escherichia coli
2.1.2.9 0.00324
-
 L-methionyl-tRNAfMet R38L mutant enzyme Escherichia coli
2.1.2.9 0.00834
-
 L-methionyl-tRNAfMet G41R/R42L double mutant enzyme Escherichia coli
2.1.2.9 0.00866
-
 L-methionyl-tRNAfMet R42K mutant enzyme Escherichia coli
2.1.2.9 0.009
-
 L-methionyl-tRNAfMet R42L mutant enzyme Escherichia coli
2.1.2.9 0.0096
-
 L-methionyl-tRNAfMet R42Q mutant enzyme Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.2.9 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O Escherichia coli involved in initiation of protein biosynthesis tetrahydrofolate + N-formylmethionyl-tRNAfMet
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.2.9 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.2.9 purification of wild-type and of R42K, R42L and G41R/R42L mutant recombinant enzymes, expressed in Escherichia coli JM109 Escherichia coli

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.2.9 additional information
-
-
 Escherichia coli

Storage Stability

EC Number Storage Stability Organism
2.1.2.9 -20°C, 20 mM imidazole, 10 mM 2-mercaptoethanol, 50% glycerol, 150 mM KCl, stable Escherichia coli
2.1.2.9 -20°C, 20 mM imidazole, pH 7.5, 10 mM 2-mercaptoethanol, 50% glycerol, 150 mM KCl, 2-4 months, stable Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.2.9 10-formyltetrahydrofolate + L-methionyl-tRNAfMet
-
 Escherichia coli tetrahydrofolate + N-formylmethionyl-tRNAfMet
-
 r
2.1.2.9 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O specificity determinants at the end of the acceptor stem of tRNAfMet Escherichia coli tetrahydrofolate + N-formylmethionyl-tRNAfMet
-
 ?
2.1.2.9 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O invariant Arg-42 within the 16-amino acid insertion loop, residues 34-49, plays a very important role both in steps of substrate binding and in catalysis, Arg-42 makes functional contacts with the determinants in the acceptor stem of the initiator tRNA Escherichia coli tetrahydrofolate + N-formylmethionyl-tRNAfMet
-
 ?
2.1.2.9 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O specificity is restricted to methionyl-tRNAfMet Escherichia coli tetrahydrofolate + N-formylmethionyl-tRNAfMet
-
 ?
2.1.2.9 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O important role of a 16-amino acid insertion loop in enzyme for tRNA recognition Escherichia coli tetrahydrofolate + N-formylmethionyl-tRNAfMet
-
 ?
2.1.2.9 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O involved in initiation of protein biosynthesis Escherichia coli tetrahydrofolate + N-formylmethionyl-tRNAfMet
-
 ?
2.1.2.9 additional information enzyme structure Escherichia coli ?
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.2.9 37
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.2.9 additional information
-
 additional information values for wild-type and mutant enzyme with wild-type and mutant initiator tRNAs Escherichia coli
2.1.2.9 0.45
-
 L-methionyl-tRNAfMet G41R/R42L double mutant enzyme Escherichia coli
2.1.2.9 0.483
-
 L-methionyl-tRNAfMet R42L mutant enzyme Escherichia coli
2.1.2.9 3.1
-
 L-methionyl-tRNAfMet R42Q mutant enzyme Escherichia coli
2.1.2.9 8.27
-
 L-methionyl-tRNAfMet R38L mutant enzyme Escherichia coli
2.1.2.9 11.4
-
 L-methionyl-tRNAfMet G41R mutant enzyme Escherichia coli
2.1.2.9 14.6
-
 L-methionyl-tRNAfMet G41K mutant enzyme Escherichia coli
2.1.2.9 27.7
-
 L-methionyl-tRNAfMet R42K mutant enzyme Escherichia coli
2.1.2.9 31.5
-
 L-methionyl-tRNAfMet G41L mutant enzyme Escherichia coli
2.1.2.9 33
-
 L-methionyl-tRNAfMet G41D mutant enzyme Escherichia coli
2.1.2.9 39.1
-
 L-methionyl-tRNAfMet wild-type enzyme Escherichia coli
2.1.2.9 46.3
-
 L-methionyl-tRNAfMet R38K mutant enzyme Escherichia coli