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Literature summary 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

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.4.33 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.4.33 0.025
-
 (Phosphate)n wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 0.055
-
 (phosphate)3 wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 0.062
-
 (Phosphate)n wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 0.11
-
 (phosphate)3 wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 0.82
-
 AMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 1.55
-
 GMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.4.33 97000
-
 gel filtration Pseudomonas aeruginosa

Organism

EC Number Organism UniProt Comment Textmining
2.7.4.33 Pseudomonas aeruginosa Q9HYF1
-
-
2.7.4.33 Pseudomonas aeruginosa DSM 22644 Q9HYF1
-
-

Substrates and Products (Substrate)

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

Subunits

EC Number Subunits Comment Organism
2.7.4.33 homodimer x-ray crystallography Pseudomonas aeruginosa

Synonyms

EC Number Synonyms Comment Organism
2.7.4.33 PA3455
-
 Pseudomonas aeruginosa
2.7.4.33 PPK2d
-
 Pseudomonas aeruginosa

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.4.33 1.9
-
 (phosphate)3 wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 3.4
-
 (phosphate)3 wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 6.7
-
 (Phosphate)n wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 13.4
-
 GMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 37.7
-
 (Phosphate)n wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 44.6
-
 AMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.4.33 8 9.5 the enzyme is active at pH 8.0-9.5 Pseudomonas aeruginosa

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.4.33 9
-
 GMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 30
-
 (phosphate)3 wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 40
-
 (phosphate)3 wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 50
-
 AMP wild type enzyme, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 270
-
 (Phosphate)n wild type enzyme, with GMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa
2.7.4.33 610
-
 (Phosphate)n wild type enzyme, with AMP as cosubstrate, at pH 8.0 temperature not specified in the publication Pseudomonas aeruginosa