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Literature summary for 2.7.4.33 extracted from

  • Nocek, B.; Kochinyan, S.; Proudfoot, M.; Brown, G.; Evdokimova, E.; Osipiuk, J.; Edwards, A.; Savchenko, A.; Joachimiaka, A.; Yakunin, A.
    Polyphosphate-dependent synthesis of ATP and ADP by the family-2 polyphosphate kinases in bacteria (2008), Proc. Natl. Acad. Sci. USA, 105, 17730-17735 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.4.33

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, using 0.1 M HEPES-Na (pH 7.0), 5% (w/v) Tacsimate, and 10% (w/v) PEG MME 5K Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
D307A the mutation leads to strongly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
D320A the mutation leads to slightly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
D323A the mutation leads to slightly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
D362A inactive Pseudomonas aeruginosa
D437A inactive Pseudomonas aeruginosa
E304A the mutation leads to strongly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
K311A the mutation leads to strongly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
K473A the mutation leads to slightly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
Q416A the mutation leads to reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
R316A the mutation leads to slightly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
R325A the mutation leads to slightly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
R363A inactive Pseudomonas aeruginosa
R419A inactive Pseudomonas aeruginosa
R423A the mutation leads to strongly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
R477A the mutation leads to slightly increased activity compared to the wild type enzyme Pseudomonas aeruginosa
W408A the mutation leads to strongly reduced activity compared to the wild type enzyme Pseudomonas aeruginosa
Y447A the mutation leads to reduced activity compared to the wild type enzyme Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.025
-
 (Phosphate)n wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
0.055
-
 (phosphate)3 wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
0.062
-
 (Phosphate)n wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
0.11
-
 (phosphate)3 wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
0.82
-
 AMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
1.55
-
 GMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required, most effective metal for activity Pseudomonas aeruginosa
additional information low activity with Co2+ or Ni2+, no activity in the presence of Mn2+ or Ca2+ Pseudomonas aeruginosa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
97000
-
 gel filtration Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9HYF1
-
-
Pseudomonas aeruginosa DSM 22644 Q9HYF1
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + (phosphate)n
-
 Pseudomonas aeruginosa AMP + (phosphate)n+1
-
 r
ADP + (phosphate)n
-
 Pseudomonas aeruginosa DSM 22644 AMP + (phosphate)n+1
-
 r
AMP + (phosphate)3
-
 Pseudomonas aeruginosa ADP + (phosphate)2
-
 r
AMP + (phosphate)3
-
 Pseudomonas aeruginosa DSM 22644 ADP + (phosphate)2
-
 r
AMP + (phosphate)n+1
-
 Pseudomonas aeruginosa ADP + (phosphate)n
-
 r
AMP + (phosphate)n+1
-
 Pseudomonas aeruginosa DSM 22644 ADP + (phosphate)n
-
 r
dAMP + (phosphate)n+1
-
 Pseudomonas aeruginosa dADP + (phosphate)n
-
 r
dAMP + (phosphate)n+1
-
 Pseudomonas aeruginosa DSM 22644 dADP + (phosphate)n
-
 r
dGMP + (phosphate)n+1
-
 Pseudomonas aeruginosa dGDP + (phosphate)n
-
 r
dGMP + (phosphate)n+1
-
 Pseudomonas aeruginosa DSM 22644 dGDP + (phosphate)n
-
 r
GDP + (phosphate)n the enzyme prefers GDP over ADP Pseudomonas aeruginosa GMP + (phosphate)n+1
-
 r
GMP + (phosphate)n+1
-
 Pseudomonas aeruginosa GDP + (phosphate)n
-
 r
IMP + (phosphate)n+1
-
 Pseudomonas aeruginosa IDP + (phosphate)n
-
 r
XMP + (phosphate)n+1
-
 Pseudomonas aeruginosa XDP + (phosphate)n
-
 r

Subunits

Subunits Comment Organism
homodimer x-ray crystallography Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
PA3455
-
 Pseudomonas aeruginosa
PPK2d
-
 Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.9
-
 (phosphate)3 wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
3.4
-
 (phosphate)3 wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
6.7
-
 (Phosphate)n wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
13.4
-
 GMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
37.7
-
 (Phosphate)n wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
44.6
-
 AMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa

pH Range

pH Minimum pH Maximum Comment Organism
8 9.5 the enzyme is active at pH 8.0-9.5 Pseudomonas aeruginosa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
9
-
 GMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
30
-
 (phosphate)3 wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
40
-
 (phosphate)3 wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
50
-
 AMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
270
-
 (Phosphate)n wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
610
-
 (Phosphate)n wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa