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Literature summary for 6.3.2.61 extracted from

  • Westermann, S.; Schneider, A.; Horn, E.K.; Weber, K.
    Isolation of tubulin polyglutamylase from Crithidia; binding to microtubules and tubulin, and glutamylation of mammalian brain alpha- and beta-tubulins (1999), J. Cell Sci., 112, 2185-2193 .
    View publication on PubMed
Search for more literature for 6.3.2.61

Activating Compound

Activating Compound Comment Organism Structure
glycerol 20% (v/v) leads to 10-15% increase in glutamylation activity Crithidia fasciculata

Inhibitors

Inhibitors Comment Organism Structure
ATP 90% decrease of activity at 10 mM ATP Crithidia fasciculata
KCl strong inhibition Crithidia fasciculata
Mg2+ 75% decrease of activity at 20 mM Mg2+ Crithidia fasciculata
NaCl about 85% inhibition at 300 mM NaCl Crithidia fasciculata

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0003
-
 [alpha/beta-tubulin]-L-glutamate at pH 9.0, temperature not specified in the publication Crithidia fasciculata
0.8
-
 L-glutamate at pH 9.0, temperature not specified in the publication Crithidia fasciculata

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoskeleton
-
 Crithidia fasciculata 5856
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ dependent on, optimal concentration of 5 mM Crithidia fasciculata

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + [alpha/beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + n L-glutamate Crithidia fasciculata (1c) [alpha/beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
ATP + [alpha/beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + L-glutamate Crithidia fasciculata (1b) [alpha/beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
ATP + [alpha/beta-tubulin]-L-glutamate + L-glutamate Crithidia fasciculata (1a) [alpha/beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
n ATP + [alpha/beta-tubulin]-L-glutamate + n L-glutamate Crithidia fasciculata overall reaction. The enzyme incorporates glutamic acid preferentially into the more acidic variants of both alpha- and beta-tubulins [alpha/beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Crithidia fasciculata
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ATP-affinity chromatography, glycerol-gradient centrifugation and Mono Q ion-exchange chromatography Crithidia fasciculata

Source Tissue

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + [alpha/beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + n L-glutamate (1c) Crithidia fasciculata [alpha/beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
ATP + [alpha/beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + L-glutamate (1b) Crithidia fasciculata [alpha/beta-tubulin]-(alpha-L-glutamyl-gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
ATP + [alpha/beta-tubulin]-L-glutamate + L-glutamate (1a) Crithidia fasciculata [alpha/beta-tubulin]-(gamma-L-glutamyl)-L-glutamate + ADP + phosphate
-
 ?
additional information synthetic peptides with an oligoglutamyl side chain, corresponding to the carboxyterminal end of brain alpha- and beta-tubulins, are accepted by the enzyme, albeit at low efficiency. The enzyme elongates the side chain by up to 3 and 5 residues, respectively. The presence of D-Glu, L-Tyr, Gly, L-Asp in 10-100fold excess over L-Glu do not affect the glutamylation activity. While mammalian brain tubulin is effectively polyglutamylated by the Crithidia enzyme, tubulin from HeLa cells is a poorer substrate Crithidia fasciculata ?
-
-
n ATP + [alpha/beta-tubulin]-L-glutamate + n L-glutamate overall reaction. The enzyme incorporates glutamic acid preferentially into the more acidic variants of both alpha- and beta-tubulins Crithidia fasciculata [alpha/beta-tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?

Subunits

Subunits Comment Organism
? x * 40000, SDS-PAGE Crithidia fasciculata

Synonyms

Synonyms Comment Organism
tubulin polyglutamylase
-
 Crithidia fasciculata

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Crithidia fasciculata

Cofactor

Cofactor Comment Organism Structure
ATP optimal concentration of 1-2 mM, cannot be replaced by GTP or ATP-gamma-S Crithidia fasciculata