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Literature summary extracted from

  • Shiba, T.; Itoh, H.; Kameda, A.; Kobayashi, K.; Kawazoe, Y.; Noguchi, T.
    Polyphosphate:AMP phosphotransferase as a polyphosphate-dependent nucleoside monophosphate kinase in Acinetobacter johnsonii 210A (2005), J. Bacteriol., 187, 1859-1865.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.4.33 gene pap, construction of a cDNA library expressed in Escherichia coli, DNA and amino acid sequence determination and analysis, overexpression of pPAP2 from plasmid in Escherichia coli strain JM109 Acinetobacter johnsonii

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.4.33 ammonium sulfate slightly inhibitory at 50 mM Acinetobacter johnsonii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.4.33 0.27
-
 AMP recombinant PAP, pH 8.0, 37°C Acinetobacter johnsonii
2.7.4.33 0.45
-
 AMP recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4 Acinetobacter johnsonii
2.7.4.33 4.4
-
 GMP recombinant PAP, pH 8.0, 37°C Acinetobacter johnsonii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.4.33 Mg2+ specifically required, lowers the Km for the nucleotide substrate Acinetobacter johnsonii
2.7.4.33 additional information other metal ions such as Mn2+, Fe2+, Ca2+, Cu2+, Zn2+, and Co2+ cannot enhance the activity Acinetobacter johnsonii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.4.33 55800
-
 x * 55800, recombinant pPAP2, SDS-PAGE Acinetobacter johnsonii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.4.33 AMP + [phosphate]n Acinetobacter johnsonii
-
 ADP + [phosphate]n-1
-
 ?
2.7.4.33 AMP + [phosphate]n Acinetobacter johnsonii 210A
-
 ADP + [phosphate]n-1
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.4.33 Acinetobacter johnsonii Q83XD3 gene pap, plasmid containing the pap gene, designated pPAP2
-
2.7.4.33 Acinetobacter johnsonii 210A Q83XD3 gene pap, plasmid containing the pap gene, designated pPAP2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.4.33 recombinant pPAP2 dimer 22.1fold from Escherichia coli strain JM109 by ammonium sulfate fractionation, anion exchange and hydroxyapatite chromatography, and gel filtration Acinetobacter johnsonii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.4.33 2.48
-
 crude extract of recombinant Escherichia coli strain JM109, pH 8.0, 37°C Acinetobacter johnsonii
2.7.4.33 24.9
-
 purified recombinant PAP tetramer, pH 8.0, 37°C Acinetobacter johnsonii
2.7.4.33 54.9
-
 purified recombinant PAP dimer, pH 8.0, 37°C Acinetobacter johnsonii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.4.33 AMP + [phosphate]n
-
 Acinetobacter johnsonii ADP + [phosphate]n-1
-
 ?
2.7.4.33 AMP + [phosphate]n best substrate Acinetobacter johnsonii ADP + [phosphate]n-1
-
 ?
2.7.4.33 AMP + [phosphate]n
-
 Acinetobacter johnsonii 210A ADP + [phosphate]n-1
-
 ?
2.7.4.33 AMP + [phosphate]n best substrate Acinetobacter johnsonii 210A ADP + [phosphate]n-1
-
 ?
2.7.4.33 CMP + [phosphate]n 0.09% of the activity with AMP Acinetobacter johnsonii CDP + [phosphate]n-1
-
 ?
2.7.4.33 CMP + [phosphate]n 0.09% of the activity with AMP Acinetobacter johnsonii 210A CDP + [phosphate]n-1
-
 ?
2.7.4.33 dAMP + [phosphate]n 18% of the activity with AMP Acinetobacter johnsonii dADP + [phosphate]n-1
-
 ?
2.7.4.33 dCMP + [phosphate]n 0.008% of the activity with AMP Acinetobacter johnsonii dCDP + [phosphate]n-1
-
 ?
2.7.4.33 dGMP + [phosphate]n 2.6% of the activity with AMP Acinetobacter johnsonii dGDP + [phosphate]n-1
-
 ?
2.7.4.33 GMP + [phosphate]n 10% of the activity with AMP Acinetobacter johnsonii GDP + [phosphate]n-1
-
 ?
2.7.4.33 GMP + [phosphate]n 10% of the activity with AMP Acinetobacter johnsonii 210A GDP + [phosphate]n-1
-
 ?
2.7.4.33 IMP + [phosphate]n 2.2% of the activity with AMP Acinetobacter johnsonii IDP + [phosphate]n-1
-
 ?
2.7.4.33 additional information PAP acts as a poly(P)-dependent NMP kinase, activity for AMP by PAP is about 10times greater than that for GMP and 770 and about 1100times greater than that for UMP and CMP. ATP production by the coupled reactions of recombinant PAP and purified Escherichia coli poly(P) kinases from AMP. The purified recombinant PAP enzyme has not only strong PAP activity but also shows poly(P)-dependent nucleoside monophosphate kinase activity, by which it converts ribonucleoside monophosphates and deoxyribonucleoside monophosphates into ribonucleoside diphosphates and deoxyribonucleoside diphosphates, respectively Acinetobacter johnsonii ?
-
 ?
2.7.4.33 additional information PAP acts as a poly(P)-dependent NMP kinase, activity for AMP by PAP is about 10times greater than that for GMP and 770 and about 1100times greater than that for UMP and CMP. ATP production by the coupled reactions of recombinant PAP and purified Escherichia coli poly(P) kinases from AMP. The purified recombinant PAP enzyme has not only strong PAP activity but also shows poly(P)-dependent nucleoside monophosphate kinase activity, by which it converts ribonucleoside monophosphates and deoxyribonucleoside monophosphates into ribonucleoside diphosphates and deoxyribonucleoside diphosphates, respectively Acinetobacter johnsonii 210A ?
-
 ?
2.7.4.33 TMP + [phosphate]n 0.012% of the activity with AMP Acinetobacter johnsonii TDP + [phosphate]n-1
-
 ?
2.7.4.33 UMP + [phosphate]n 0.13% of the activity with AMP Acinetobacter johnsonii UDP + [phosphate]n-1
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.4.33 ? x * 55800, recombinant pPAP2, SDS-PAGE Acinetobacter johnsonii

Synonyms

EC Number Synonyms Comment Organism
2.7.4.33 PAP
-
 Acinetobacter johnsonii
2.7.4.33 polyphosphate:AMP phosphotransferase
-
 Acinetobacter johnsonii
2.7.4.33 pPAP2
-
 Acinetobacter johnsonii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.4.33 50
-
 recombinant enzyme Acinetobacter johnsonii

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.4.33 30 50 the level of activity at 50°C is 3.2fold and 1.8fold higher than that at 30°C and 37°C, respectively Acinetobacter johnsonii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.4.33 45
-
 purified recombinant PAP, 10 min, 40% activity remaining without polyphosphate, 85% activity remaining in presence of 10 mM polyphosphate Acinetobacter johnsonii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.4.33 35
-
 GMP recombinant PAP, pH 8.0, 37°C Acinetobacter johnsonii
2.7.4.33 151.7
-
 AMP recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4 Acinetobacter johnsonii
2.7.4.33 166.7
-
 AMP recombinant PAP, pH 8.0, 37°C Acinetobacter johnsonii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.4.33 8 9 recombinant enzyme Acinetobacter johnsonii

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.7.4.33 7 11 purified recombinant enzyme, stable at Acinetobacter johnsonii

General Information

EC Number General Information Comment Organism
2.7.4.33 additional information PAP contains two putative phosphate-binding motifs, P-loops. The similarity between PAP and the PPK2 homologues indicates that there is a common mechanism by which these enzymes use poly(P) for phosphorylation of nucleosides. The C-terminal region of PAP, including one P-loop (amino acids 286 to 292), is thought to be essential for poly(P)-dependent nucleotide phosphorylation. An additional P-loop in the N-terminal region (amino acids 45 to 51) of PAP may play a crucial role in binding with nucleoside monophosphate Acinetobacter johnsonii

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 7.83
-
 GMP recombinant PAP, pH 8.0, 37°C Acinetobacter johnsonii
2.7.4.33 333.3
-
 AMP recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4 Acinetobacter johnsonii
2.7.4.33 616.7
-
 AMP recombinant PAP, pH 8.0, 37°C Acinetobacter johnsonii