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Literature summary extracted from

  • Regnard, C.; Audebert, S.; Desbruyeres, E.; Denoulet, P.; Edde, B.
    Tubulin polyglutamylase partial purification and enzymatic properties (1998), Biochemistry, 37, 8395-8404 .
    View publication on PubMed

General Stability

EC Number General Stability Organism
6.3.2.61 at 30°C, the loss of activity is reduced to 40% in the presence of 0.1 mg/ml microtubules, while neither bovine serum albumin nor the other substrates, ATP or glutamate, have any protecting effect Mus musculus

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.2.61 citrate about 90% inhibition at 10 mM Mus musculus
6.3.2.61 KCl 5-10 mM, strong inhibitory effect on the glutamylation reaction. 80 mM, 90% inhibition; strong inhibitory effect Mus musculus
6.3.2.61 Na2SO4 90% inhibition at 20 mM Mus musculus
6.3.2.61 NaCl 5-10 mM, strong inhibitory effect on the glutamylation reaction. 40 mM NaCl, 50% inhibition; strong inhibitory effect Mus musculus
6.3.2.61 Sodium acetate 100 mM, 50% onhibition Mus musculus
6.3.2.61 Sodium citrate 90% inhibition at 10 mM Mus musculus
6.3.2.61 Sodium sulfate about 90% inhibition at 20 mM Mus musculus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.61 0.1
-
 ATP 30°C, pH 8.7 Mus musculus
6.3.2.61 0.8
-
 L-glutamate 30°C, pH 8.7 Mus musculus
6.3.2.61 3
-
 L-glutamate 30°C, pH 6.8 Mus musculus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.2.61 Mg2+ absolutely required with an optimal concentration of 6-8 mM Mus musculus
6.3.2.61 Mg2+ required, 8 mM used in assay conditions. The activity increases markedly with Mg2+ concentrations between 1 and 6 mM Mus musculus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.2.61 360000
-
-
 Mus musculus
6.3.2.61 360000
-
 gel filtration Mus musculus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.61 n ATP + [alpha-tubulin]-L-glutamate + n L-glutamate Mus musculus overall reaction [alpha-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
6.3.2.61 n ATP + [beta-tubulin]-L-glutamate + n L-glutamate Mus musculus overall reaction [beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.61 Mus musculus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.61
-
 Mus musculus
6.3.2.61 phosphocellulose column chromatography, MgCl2 and sodium citrate precipitation, sucrose gradient sedimentation, and TSK G3000 gel filtration Mus musculus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
6.3.2.61 brain
-
 Mus musculus
-

Storage Stability

EC Number Storage Stability Organism
6.3.2.61 -80°C, 10% glycerol, stable for several months Mus musculus
6.3.2.61 -80°C, in the presence of 10% (v/v) glycerol, several months, no loss of activity Mus musculus
6.3.2.61 20°C, in 10 mM Tris (pH 8.7), 1 mM EGTA, 1 mM MgCl2, 1 mM dithiothreitol, and 0.01% TX-100 at pH 8.7, 1 h, 50% loss of activity Mus musculus
6.3.2.61 30°C, in 10 mM Tris (pH 8.7), 1 mM EGTA, 1 mM MgCl2, 1 mM dithiothreitol, and 0.01% TX-100 at pH 8.7, 1 h, 75% loss of activity Mus musculus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.61 ATP + [alpha-tubulin]-L-glutamate + L-glutamate the enzyme catalyzes the MgATP-dependent addition of L-glutamate onto tubulin subunits. The enzyme is active with alpha- and beta-tubulin, respectively. Microtubules are much better substrates than unpolymerized tubulin, and the reaction is very specific for glutamate, other amino acids or glutamate analogues not being substrates. Moreover, glutamyl units are added sequentially onto tubulin, leading to progressive elongation of the polyglutamyl side chains. Side chains of one to six or seven glutamyl units are obtained with microtubules, whereas much longer side chains (up to 15-20 units) are formed with unpolymerized tubulin Mus musculus ?
-
 ?
6.3.2.61 ATP + [beta-tubulin]-L-glutamate + L-glutamate the enzyme catalyzes the MgATP-dependent addition of L-glutamate onto tubulin subunits. The enzyme is active with alpha- and beta-tubulin, respectively. Microtubules are much better substrates than unpolymerized tubulin, and the reaction is very specific for glutamate, other amino acids or glutamate analogues not being substrates. Moreover, glutamyl units are added sequentially onto tubulin, leading to progressive elongation of the polyglutamyl side chains. Side chains of one to six or seven glutamyl units are obtained with microtubules, whereas much longer side chains (up to 15-20 units) are formed with unpolymerized tubulin Mus musculus ?
-
 ?
6.3.2.61 additional information microtubules are much better substrates than unpolymerized tubulin, and the reaction is very specific for L-glutamate, other amino acids (D-Glu, L-Asp, L-Gln, L-Asn, L-Ser, L-Lys, L-Leu, and L-Gly) or glutamate analogues (RGlu-Glu, çGlu-Glu, Iso-Gln, N-acetyl-Glu, norvaline, GABA, and Gla) not being substrates. Moreover, glutamyl units are added sequentially onto tubulin, leading to progressive elongation of the polyglutamyl side chains. Side chains of 1-6 or 7 glutamyl units are obtained with microtubules, whereas much longer side chains (up to 15-20 units) are formed with unpolymerized tubulin Mus musculus ?
-
-
6.3.2.61 n ATP + [alpha-tubulin]-L-glutamate + n L-glutamate overall reaction Mus musculus [alpha-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
6.3.2.61 n ATP + [beta-tubulin]-L-glutamate + n L-glutamate overall reaction Mus musculus [beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.2.61 tubulin polyglutamylase
-
 Mus musculus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
6.3.2.61 20
-
 pH 8.7, 1 h, 50% loss of activity. Not significant loss of activity within 3 h of incubation at pH 6.8 Mus musculus
6.3.2.61 30
-
 pH 8.7, 1 h, 75% loss of activity. No significant loss of activity within 3 h of incubation at pH 6.8 Mus musculus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.61 additional information
-
 the high pH optimum observed at low glutamate concentrations is a result of the free amine of glutamate (pK = 9.7 at 25°C) being the substrate for the reaction Mus musculus
6.3.2.61 8.3 8.9
-
 Mus musculus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
6.3.2.61 6.8 8.9 the enzyme activity increases markedly with the pH going from 6.8 to 8.3, with a large optimum between 8.3 and 8.9, and then decreases sharply. At physiological pH, the activity is 15% of the maximal level Mus musculus
6.3.2.61 7.5 9.2 pH 7.5: about 45% of maximal activity, pH 9.2: about 80% of maximal activity. When the glutamate concentration is raised to 6 mM, the activity is less dependent on pH, decreasing by only 40% between pH 8.7 and 6.8 Mus musculus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
6.3.2.61 6.8
-
 3 h, no significant loss of activity at 20°C and at 30°C Mus musculus
6.3.2.61 8.7
-
 1 h, 50% loss of activity at 20°C and 75% at 30°C. At 30°C, the loss is reduced to 40% in the presence of 0.1 mg/ml microtubules Mus musculus

General Information

EC Number General Information Comment Organism
6.3.2.61 physiological function the enzyme catalyzes the posttranslational formation of polyglutamyl side chains onto alpha- and beta-tubulin. The length of the polyglutamyl side chain regulates the interaction between tubulin and various microtubule-associated proteins Mus musculus