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Literature summary for 6.3.2.61 extracted from

  • Mahalingan, K.K.; Keith Keenan, E.; Strickland, M.; Li, Y.; Liu, Y.; Ball, H.L.; Tanner, M.E.; Tjandra, N.; Roll-Mecak, A.
    Structural basis for polyglutamate chain initiation and elongation by TTLL family enzymes (2020), Nat. Struct. Mol. Biol., 27, 802-813.
    View publication on PubMed
Search for more literature for 6.3.2.61

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Arctic Express (DE3) cells Mus musculus

Crystallization (Commentary)

Crystallization (Comment) Organism
wild type and mutant enzyme C179A/Q180R/H362I, in complex with ATP and inhibitors, hanging dop vapor diffusion method, using 0.2 M ammonium sulfate and 10-15% (w/v) PEG 3350 Mus musculus

Protein Variants

Protein Variants Comment Organism
C179A/Q180R/H362I the mutant no longer discriminates between heterogenous glutamylated brain microtubules and unmodified microtubules, as both the kcat and Km are similar with these substrates. The triple-mutant has strong preference for glutamylating an unmodified alpha1B peptide Mus musculus
K283A the mutation reduces activity to the background level Mus musculus
K377A the mutation reduces activity to the background level Mus musculus
Q180A he mutation decreases the enzyme activity by 64% compared to the wild type enzyme Mus musculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00773
-
 [unmodified human tubulin]-L-glutamate pH and temperature not specified in the publication Mus musculus
0.00809
-
 [modified porcine brain tubulin]-L-glutamate pH and temperature not specified in the publication Mus musculus

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Mus musculus A4Q9E8
-
-

Purification (Commentary)

Purification (Comment) Organism
GST affinity chromatography, heparin column chromatography, and Superdex 75 gel filtration Mus musculus

Source Tissue

Source Tissue Comment Organism Textmining
brain
-
 Mus musculus
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information isoform TTLL6 has barely any activity on unmodified recombinant human alpha1A/betaIII microtubules, but about 20fold higher activity on detyrosinated alpha1A-Y/betaIII microtubules where the alpha-tail has a terminal glutamate with an available alpha-carboxyl group Mus musculus ?
-
-
n ATP + [modified porcine brain tubulin]-L-glutamate + n L-glutamate
-
 Mus musculus [modified porcine brain tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
n ATP + [tubulin]-L-glutamate + n L-glutamate
-
 Mus musculus [tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?
n ATP + [unmodified human tubulin]-L-glutamate + n L-glutamate
-
 Mus musculus [unmodified human tubulin]-(gamma-(poly-alpha-L-glutamyl)-L-glutamyl)-L-glutamate + n ADP + n phosphate
-
 ?

Synonyms

Synonyms Comment Organism
ttll6
-
 Mus musculus
tubulin tyrosine ligase-like
-
 Mus musculus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0032
-
 [modified porcine brain tubulin]-L-glutamate pH and temperature not specified in the publication Mus musculus
0.0037
-
 [unmodified human tubulin]-L-glutamate pH and temperature not specified in the publication Mus musculus