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Literature summary extracted from

  • Mukai, M.; Ikegami, K.; Sugiura, Y.; Takeshita, K.; Nakagawa, A.
    Mitsutoshi Setou Recombinant mammalian tubulin polyglutamylase TTLL7 performs both initiation and elongation of polyglutamylation on - tubulin through a random sequential pathway (2009), Biochemistry, 48, 1084-1093 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.62 expressed in Escherichia coli Rosetta (DE3) cells Mus musculus

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.2.62 Adenylyl imidodiphosphate pH 7.0, 30°C, at a 600fold higher concentration compared with the ATP concentration, almost completely inhibits the enzyme. Competitive inhibition for ATP. Noncompetitive inhibitor for glutamate and tubulin Mus musculus
6.3.2.62 ADP about 98% inhibition at 30 mM; pH 7.0, 30°C, at a 600fold higher concentration compared with the ATP concentration, almost completely inhibits the enzyme Mus musculus
6.3.2.62 AMP about 98% inhibition at 30 mM; pH 7.0, 30°C, at a 600fold higher concentration compared with the ATP concentration, almost completely inhibits the enzyme Mus musculus
6.3.2.62 AMP-PNP about 98% inhibition at 30 mM, competitive inhibitor for ATP, noncompetitive inhibitorn for glutamate and tubulin Mus musculus
6.3.2.62 ATAEEEGEMYEDDDEESEAQGPK about 80% inhibition at 10 mM Mus musculus
6.3.2.62 C-terminal peptide of class III-tubulin competitive inhibitor for tubulin. Noncompetitive inhibition against glutamate and ATP Mus musculus
6.3.2.62 DATAEEGEFEEEAEEEVA L about 70% inhibition at 10 mM Mus musculus
6.3.2.62 gamma-L-glutamyl-L-cysteine about 58% inhibition at 10 mM Mus musculus
6.3.2.62 gamma-L-glutamyl-L-glutamate pH 7.0, 30°C, competitive inhibitor for glutamate. Noncompetitive inhibitor for ATP and tubulin Mus musculus
6.3.2.62 gamma-L-glutamyl-L-leucine about 60% inhibition at 10 mM Mus musculus
6.3.2.62 gammaGlu-Glu 80% inhibition at 10 mM, noncompetitive inhibitor for ATP and tubulin Mus musculus
6.3.2.62 Gly less than 10% inhibition at 10 mM Mus musculus
6.3.2.62 L-Asp about 35% inhibition at 10 mM Mus musculus
6.3.2.62 L-Gln about 50% inhibition at 10 mM Mus musculus
6.3.2.62 L-His about 40% inhibition at 10 mM Mus musculus
6.3.2.62 L-Leu about 15% inhibition at 10 mM Mus musculus
6.3.2.62 L-Tyr about 10% inhibition at 10 mM Mus musculus
6.3.2.62 NaCl the polyglutamylase activity of the recombinant enzyme is inhibited by NaCl in proportion to the concentrations of NaCl: 100 mM NaCl suppresses the enzyme activity at 40%, and 200 mM NaCl suppressed it at 90% Mus musculus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.62 0.0125
-
 [beta-tubulin]-L-glutamate at pH 7.0 and 30°C Mus musculus
6.3.2.62 0.0128
-
 L-glutamate at pH 7.0 and 30°C Mus musculus
6.3.2.62 0.0133
-
 ATP at pH 7.0 and 30°C Mus musculus
6.3.2.62 3.1
-
 [beta-tubulin]-L-glutamate pH 7.0, 30°C, enzyme from adult brain, 0.006 mM L-glutamate Mus musculus
6.3.2.62 4.4
-
 L-glutamate pH 7.0, 30°C, enzyme from newborn brain, 0.05 mM ATP Mus musculus
6.3.2.62 5.4
-
 L-glutamate pH 7.0, 30°C, enzyme from adult brain, 0.05 mM ATP Mus musculus
6.3.2.62 6.7
-
 [beta-tubulin]-L-glutamate pH 7.0, 30°C, enzyme from newborn brain, 0.05 mM L-glutamate Mus musculus
6.3.2.62 7.5
-
 ATP pH 7.0, 30°C, enzyme from adult brain, 0.006 mM L-glutamate Mus musculus
6.3.2.62 15.8
-
 [beta-tubulin]-L-glutamate pH 7.0, 30°C, enzyme from newborn brain, 0.05 mM ATP Mus musculus
6.3.2.62 16
-
 ATP pH 7.0, 30°C, enzyme from newborn brain, 0.05 mM L-glutamate Mus musculus
6.3.2.62 19.1
-
 ATP pH 7.0, 30°C, enzyme from adult brain, 0.017 mM tubulin Mus musculus
6.3.2.62 19.6
-
 L-glutamate pH 7.0, 30°C, enzyme from newborn brain, 0.017 mM tubulin Mus musculus
6.3.2.62 20.2
-
 L-glutamate pH 7.0, 30°C, enzyme from adult brain, 0.017 mM tubulin Mus musculus
6.3.2.62 21.8
-
 [beta-tubulin]-L-glutamate pH 7.0, 30°C, enzyme from adult brain, 0.05 mM ATP Mus musculus
6.3.2.62 26.5
-
 ATP pH 7.0, 30°C, enzyme from newborn brain, 0.017 mM tubulin Mus musculus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.2.62 Mg2+ 8 mM used in assay conditions Mus musculus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.62 ATP + [beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + n L-glutamate Mus musculus (1c) [beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
6.3.2.62 ATP + [beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + L-glutamate Mus musculus (1b) [beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
6.3.2.62 ATP + [beta-tubulin]-L-glutamate + L-glutamate Mus musculus (1a) [beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
6.3.2.62 n ATP + [beta-tubulin]-L-glutamate + n L-glutamate Mus musculus the whole activity to microtubules is about 2.5fold higher than that to unpolymerized tubulin. When microtubules are used as substrates, recombinant enzyme exhibits activity highly preferential to beta-tubulin. The substrate preference drastically decreases when unpolymerized tubulin dimers are used as substrates [beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.62 Mus musculus
-
-
-
6.3.2.62 Mus musculus A4Q9F0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.62 glutathione-Sepharose bead chromatography, and Superdex 200 gel filtration Mus musculus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
6.3.2.62 brain adult and newborn Mus musculus
-

Storage Stability

EC Number Storage Stability Organism
6.3.2.62 -80°C, 50 mM Tris-HCl (pH 7.4), 20 mM NaCl, 1 mM EDTA, and 1 mM dithiothreitol, at least 3 months, no loss of activity Mus musculus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.62 ATP + [beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + n L-glutamate (1c) Mus musculus [beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
6.3.2.62 ATP + [beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + L-glutamate (1b) Mus musculus [beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
6.3.2.62 ATP + [beta-tubulin]-L-glutamate + L-glutamate (1a) Mus musculus [beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
6.3.2.62 ATP + [beta-tubulin]-L-glutamate + L-glutamate the enzyme performs both initiation and elongation in the polyglutamylation reaction on beta-tubulin through a random sequential pathway Mus musculus [tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
6.3.2.62 n ATP + [beta-tubulin]-L-glutamate + n L-glutamate the whole activity to microtubules is about 2.5fold higher than that to unpolymerized tubulin. When microtubules are used as substrates, recombinant enzyme exhibits activity highly preferential to beta-tubulin. The substrate preference drastically decreases when unpolymerized tubulin dimers are used as substrates Mus musculus [beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.2.62 beta-tubulin-selective polyglutamylase
-
 Mus musculus
6.3.2.62 TTLL7
-
 Mus musculus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.62 7
-
-
 Mus musculus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.2.62 0.02
-
 Adenylyl imidodiphosphate pH 7.0, 30°C, versus ATP Mus musculus
6.3.2.62 0.12
-
 Adenylyl imidodiphosphate pH 7.0, 30°C, versus L-glutamate Mus musculus
6.3.2.62 0.23
-
 Adenylyl imidodiphosphate pH 7.0, 30°C, versus beta-tubulin Mus musculus
6.3.2.62 0.5
-
 C-terminal peptide of class III-tubulin pH 7.0, 30°C, versus beta-tubulin Mus musculus
6.3.2.62 0.83
-
 C-terminal peptide of class III-tubulin pH 7.0, 30°C, versus ATP Mus musculus
6.3.2.62 1.06
-
 C-terminal peptide of class III-tubulin pH 7.0, 30°C, versus L-glutamate Mus musculus

General Information

EC Number General Information Comment Organism
6.3.2.62 metabolism the enzyme TTLL7 performs both initiation and elongation in the polyglutamylation reaction on beta-tubulin Mus musculus