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Literature summary for 2.7.3.3 extracted from

  • Voncken, F.; Gao, F.; Wadforth, C.; Harley, M.; Colasante, C.
    The phosphoarginine energy-buffering system of Trypanosoma brucei involves multiple arginine kinase isoforms with different subcellular locations (2013), PLoS ONE, 8, e65908.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene Tb09.160.4560, sequence comparisons of Trypanosoma brucei isozymes AK1-3, recombinant expression of N-terminally His-tagged isozyme in Escherichia coli strain BL21(DE3) Trypanosoma brucei brucei
gene Tb927.9.6230, sequence comparisons of Trypanosoma brucei isozymes AK1-3 Trypanosoma brucei brucei
gene TbAK3, sequence comparisons of Trypanosoma brucei isozymes AK1-3 Trypanosoma brucei brucei

Protein Variants

Protein Variants Comment Organism
additional information generation of tetracycline-inducible PCF TbAK-RNAi cell line which allows the simultaneous silencing of all three TbAK isoforms. The TbAK-depleted cell line becomes about 400fold more sensitive to hydrogen peroxide Trypanosoma brucei brucei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.23
-
ATP pH 7.5, 28°C, recombinant isozyme AK1 Trypanosoma brucei brucei
0.24
-
L-arginine pH 7.5, 28°C, recombinant isozyme AK1 Trypanosoma brucei brucei
0.32
-
ATP pH 7.5, 37°C, recombinant isozyme AK1 Trypanosoma brucei brucei
0.48
-
L-arginine pH 7.5, 37°C, recombinant isozyme AK1 Trypanosoma brucei brucei

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol isozyme TbAK3 is exclusively found in the cytosol. Isozyme TbAK3 lacks any apparent targeting sequences and is accordingly located in the cytosol of the parasite Trypanosoma brucei brucei 5829
-
flagellum isozyme TbAK1 is exclusively found in the flagellum. The flagellar location of TbAK1 is dependent on a 22 amino acid long N-terminal sequence, which is sufficient for targeting a GFP-fusion protein to the trypanosome flagellum Trypanosoma brucei brucei
-
-
glycosome isozyme TbAK2 is exclusively found in the glycosome. The glycosomal location of TbAK2 is in agreement with the presence of a conserved peroxisomal targeting signal, the C-terminal tripeptide SNL Trypanosoma brucei brucei 20015
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Trypanosoma brucei brucei

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
-
Trypanosoma brucei brucei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-arginine Trypanosoma brucei brucei
-
ADP + Nomega-phospho-L-arginine
-
r
ATP + L-arginine Trypanosoma brucei brucei 927/4
-
ADP + Nomega-phospho-L-arginine
-
r
ATP + L-arginine Trypanosoma brucei brucei 927 / 4 GUTat10.1 / TREU927
-
ADP + Nomega-phospho-L-arginine
-
r

Organism

Organism UniProt Comment Textmining
Trypanosoma brucei brucei Q38EU9 gene TbAK3 or Tb927.9.6290
-
Trypanosoma brucei brucei Q38EV1 gene Tb927.9.6230
-
Trypanosoma brucei brucei Q38EV2
-
-
Trypanosoma brucei brucei 927 / 4 GUTat10.1 / TREU927 Q38EV2
-
-
Trypanosoma brucei brucei 927/4 Q38EU9 gene TbAK3 or Tb927.9.6290
-
Trypanosoma brucei brucei 927/4 Q38EV1 gene Tb927.9.6230
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography Trypanosoma brucei brucei

Source Tissue

Source Tissue Comment Organism Textmining
additional information each TbAK isoform is differentially expressed in bloodstream and procyclic forms of Trypanosoma brucei, while the total cellular arginine kinase activity is 3fold higher in bloodstream form trypanosomes Trypanosoma brucei brucei
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-arginine
-
Trypanosoma brucei brucei ADP + Nomega-phospho-L-arginine
-
r
ATP + L-arginine
-
Trypanosoma brucei brucei 927/4 ADP + Nomega-phospho-L-arginine
-
r
ATP + L-arginine
-
Trypanosoma brucei brucei 927 / 4 GUTat10.1 / TREU927 ADP + Nomega-phospho-L-arginine
-
r

Subunits

Subunits Comment Organism
? x * 55000, recombinant His-tagged isozyme AK1, SDS-PAGE Trypanosoma brucei brucei

Synonyms

Synonyms Comment Organism
Tb09.160.4560
-
Trypanosoma brucei brucei
TbAK2
-
Trypanosoma brucei brucei
TbAK3
-
Trypanosoma brucei brucei

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
28 37 assay at Trypanosoma brucei brucei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Trypanosoma brucei brucei

Cofactor

Cofactor Comment Organism Structure
ATP
-
Trypanosoma brucei brucei

General Information

General Information Comment Organism
malfunction elimination of the total cellular arginine kinase activity by RNA interference significantly decreases growth of procyclic form Trypanosoma brucei by 90% under standard culture conditions and is lethal for this life cycle stage in the presence of hydrogen peroxide Trypanosoma brucei brucei
physiological function the phosphoarginine energy-buffering system of Trypanosoma brucei involves multiple arginine kinase isoforms with different subcellular locations. Increased arginine kinase activity improves growth of procyclic form Trypanosoma brucei during oxidative challenges with hydrogen peroxide Trypanosoma brucei brucei