Cloned (Comment) | Organism |
---|---|
gene AK1, DNA and amino acid sequence determination and analysis, recombinant expression of C-terminally His6-tagged isozyme AK1 in Escherichia coli strain BL21 (DE3). The prokaryotic protein expression host Escherichia coli uses the standard genetic code while Tetrahymena pyriformis uses an alternative genetic code where TAA and TAG code for glutamine instead of a stop codon, therefore a TAA codon in AK1 is replaced with CAA | Tetrahymena pyriformis |
gene AK2, DNA and amino acid sequence determination and analysis, recombinant expression of C-terminally His6-tagged isozyme AK2 in Escherichia coli strain BL21(DE3). The prokaryotic protein expression host Escherichia coli uses the standard genetic code while Tetrahymena pyriformis uses an alternative genetic code where TAA and TAG code for glutamine instead of a stop codon., therefore two TAA and three TAG codons in AK2 are replaced with CAA and CAG | Tetrahymena pyriformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetic analysis, overview. The enzyme shows a random-order, rapid equilibrium kinetic mechanism | Tetrahymena pyriformis | |
0.26 | - |
L-arginine | pH 8.0, 25°C, recombinant His6-tagged isozyme AK1 | Tetrahymena pyriformis | |
0.35 | - |
L-arginine | pH 8.0, 25°C, recombinant His6-tagged isozyme AK2 | Tetrahymena pyriformis | |
0.73 | - |
ATP | pH 8.0, 25°C, recombinant His6-tagged isozyme AK1 | Tetrahymena pyriformis | |
0.73 | - |
ATP | pH 8.0, 25°C, recombinant His6-tagged isozyme AK2 | Tetrahymena pyriformis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytoplasm | - |
Tetrahymena pyriformis | 5737 | - |
membrane | the amino acid sequence of Tetrahymena pyriformis isozyme AK1, but not isozyme AK2, has a distinct myristoylation signal sequence at the N-terminus, suggesting that 40-kDa AK1 targets the membrane | Tetrahymena pyriformis | 16020 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Tetrahymena pyriformis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
- |
Tetrahymena pyriformis |
80000 | - |
- |
Tetrahymena pyriformis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-arginine | Tetrahymena pyriformis | - |
ADP + Nomega-phospho-L-arginine | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Tetrahymena pyriformis | U3T1K4 | isozyme AK1 | - |
Tetrahymena pyriformis | U3T2Y2 | isozyme AK2 | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
lipoprotein | isozyme AK1 has a distinct myristoylation signal sequence at the N-terminus | Tetrahymena pyriformis |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged isozyme AK1 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography | Tetrahymena pyriformis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + L-arginine = ADP + Nomega-phospho-L-arginine | random-order, rapid equilibrium kinetic mechanism | Tetrahymena pyriformis |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
flagellum | - |
Tetrahymena pyriformis | - |
additional information | isozyme AK1 is mainly localized in the ciliary fraction, AK1 is attached to the ciliary membrane through the myristoylation group | Tetrahymena pyriformis | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-arginine | - |
Tetrahymena pyriformis | ADP + Nomega-phospho-L-arginine | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 40000, isozyme AK1, SDS-PAGE | Tetrahymena pyriformis |
? | x * 80000, isozyme AK2, SDS-PAGE | Tetrahymena pyriformis |
Synonyms | Comment | Organism |
---|---|---|
AK1 | - |
Tetrahymena pyriformis |
AK2 | - |
Tetrahymena pyriformis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Tetrahymena pyriformis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
63.4 | - |
ATP | pH 8.0, 25°C, recombinant His6-tagged isozyme AK2 | Tetrahymena pyriformis | |
63.4 | - |
L-arginine | pH 8.0, 25°C, recombinant His6-tagged isozyme AK2 | Tetrahymena pyriformis | |
104 | - |
ATP | pH 8.0, 25°C, recombinant His6-tagged isozyme AK1 | Tetrahymena pyriformis | |
104 | - |
L-arginine | pH 8.0, 25°C, recombinant His6-tagged isozyme AK1 | Tetrahymena pyriformis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Tetrahymena pyriformis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ADP | - |
Tetrahymena pyriformis | |
ATP | - |
Tetrahymena pyriformis |
General Information | Comment | Organism |
---|---|---|
evolution | two putative AK genes in the genome of Tetrahymena thermophila: one is a typical AK with a 40-kDa subunit (AK1) and the other is an unusual two-domain AK2 having an 80-kDa contiguous dimer, which appears to be the result of gene duplication and subsequent fusion | Tetrahymena pyriformis |
additional information | the five residues S63, Y68, E225, C271, and E314 that interact with the arginine substrate are conserved, as well as the five Arg residues R124, R126, R229, R280 and R309 that interact with substrate ATP. Residues D62 and R193 are suggested to play a key role in stabilizing the substrate-bound structures of AK by forming an ion pair. Tyr89 is also a key residue in typical invertebrate AKs and is strictly conserved. This residue is not directly involved in substrate binding but it is located close to the site that bindswith the substrate arginine, it significantly and specifically affects guanidino substrate | Tetrahymena pyriformis |
physiological function | isozyme AK1 plays a role in the phosphoarginine shuttle, which enables a continuous energy flow to dynein for ciliary movement in Tetrahymena pyriformis | Tetrahymena pyriformis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
86.9 | - |
ATP | pH 8.0, 25°C, recombinant His6-tagged isozyme AK2 | Tetrahymena pyriformis | |
142.5 | - |
ATP | pH 8.0, 25°C, recombinant His6-tagged isozyme AK1 | Tetrahymena pyriformis | |
181.1 | - |
L-arginine | pH 8.0, 25°C, recombinant His6-tagged isozyme AK2 | Tetrahymena pyriformis | |
400 | - |
L-arginine | pH 8.0, 25°C, recombinant His6-tagged isozyme AK1 | Tetrahymena pyriformis |