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Literature summary for 3.6.5.2 extracted from

  • Francis, J.W.; Newman, L.E.; Cunningham, L.A.; Kahn, R.A.
    A trimer consisting of the tubulin-specific chaperone D (TBCD), regulatory GTPase ARL2, and beta-tubulin is required for maintaining the microtubule network (2017), J. Biol. Chem., 292, 4336-4349 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.6.5.2

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of HA- and GST-tagged wild-type and mutant enzymes in HEK cells, recombinant optimized expression of Arl2 in HeLa cells Homo sapiens

Protein Variants

Protein Variants Comment Organism
F50A site-directed mutagenesis, the mutation disrupts the interaction with TBCD, increased expression levels compared to wild-type Homo sapiens
F50A/Q70L site-directed mutagenesis, cells expressing mutant L3A/F50A show a reversal of the microtubule loss phenotype, with about 20% of transfected cells showing only intermediate loss in microtubule densities Homo sapiens
I6R site-directed mutagenesis, the mutation disrupts the interaction with TBCD Homo sapiens
I6R/Q70L site-directed mutagenesis, the addition of the I6R mutation to Q70L does not reverse the effects of the dominant mutant on microtubule densities Homo sapiens
L3A site-directed mutagenesis, the mutation disrupts the interaction with TBCD Homo sapiens
L3A/Q70L site-directed mutagenesis, cells expressing mutant L3A/Q70L show a reversal of the microtubule loss phenotype, with about 10% of transfected cells showing only intermediate loss in microtubule densities Homo sapiens
additional information construction of truncaction mutant DELTA1-9. All Arl2 mutants co-purify to similar levels as wild-type ARL2 with TBCD and tubulin in a complex, with no apparent differences in the composition of the trimer Homo sapiens
Q70L site-directed mutagenesis, ARL2 dominant activating mutation. Expression of ARL2 Q70L mutant causes the loss of polymerized microtubules in cultured cells Homo sapiens
T30N site-directed mutagenesis, ARL2 dominant inactivating mutation, shows decreased expression levels compared to wild-type Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
 when human ARL2 is expressed in bacteria, it purifies as a soluble, stable monomer with no evidence of any species greater than 20 kDa Homo sapiens
20000
-
 recombinant Arl2, gel filtration Homo sapiens
200000
-
 Arl2-TBCD-tubulin complex, gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
GTP + H2O Homo sapiens
-
 GDP + phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens P36404
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged wild-type and mutant enzymes from HEK cells by glutathione affinity chromatography. Purification GST-TBCD with or withoout ARL2 from HEK cells results in the co-purification of different combinations of tubulins in novel complexes Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
brain
-
 Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GTP + H2O
-
 Homo sapiens GDP + phosphate
-
 ?

Synonyms

Synonyms Comment Organism
ADP-ribosylation factor-like protein 2 UniProt Homo sapiens
ARL2
-
 Homo sapiens
small GTPase
-
 Homo sapiens

General Information

General Information Comment Organism
additional information GST-TBCD with or without ARL2 from HEK cells co-purifies with different combinations of tubulins in novel complexes. The TBCD-alpha-tubulin-beta-tubulin trimer is analyzed, overview Homo sapiens
physiological function ARL2 plays an essential role in the cellular actions of tubulin binding cofactor B (TBCD) but, rather than competing with the binding of beta-tubulin, facilitates it. The ARL2-TBCD interaction is critical for proper maintenance of microtubule densities in cells. The TBCD-ARL2-beta-tubulin trimer represents a functional complex whose activity is fundamental to microtubule dynamics. Correlation between the ability of ARL2 and ARL2 mutants to bind TBCD and to alter microtubule dynamics in a cell-based assay Homo sapiens