Q58327 (NPPNK_METJA) Reviewed, UniProtKB/Swiss-Prot
Last modified July 9, 2014. Version 92. History...
Names and origin
|Protein names||Recommended name:|
Bifunctional NADP phosphatase/NAD kinase
|Organism||Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440) (Methanococcus jannaschii) [Reference proteome] [HAMAP]|
|Taxonomic identifier||243232 [NCBI]|
|Taxonomic lineage||Archaea › Euryarchaeota › Methanococci › Methanococcales › Methanocaldococcaceae › Methanocaldococcus ›|
|Sequence length||574 AA.|
|Protein existence||Evidence at protein level|
General annotation (Comments)
Involved in the regulation of the intracellular balance between NAD(H) and NADP(H), and is a key enzyme in the biosynthesis of NADP. Catalyzes the phosphorylation and dephosphorylation of NAD and NADP, respectively. Although it shows conflicting dual activities and is able to supply NADP, it seems that its physiological role is to prevent excess accumulation of NADP. Kinase can use ATP and other nucleoside triphosphates (UTP, TTP, CTP, GTP) as well as inorganic polyphosphate (poly(P)) as phosphoryl donors, however poly(P) is not considered to be the physiological phosphoryl donor. NAD is the preferred substrate for the kinase, but NADH can also be used as phosphoryl acceptor. Phosphatase can use NADP or NADPH as phosphoryl donor, but NADP is the preferred substrate. Phosphatase also has an activity toward the terminal phosphate group at C-2 of adenosine in 2'-AMP and toward the phosphate group at C-1 of fructose 1,6-bisphosphate, but not toward inositol 1-phosphate. Ref.2
ATP + NAD+ = ADP + NADP+. Ref.2
NADP+ + H2O = NAD+ + phosphate. Ref.2
Magnesium. NAD and pNPPase kinase show maximum activities at 50 and 20mM magnesium, respectively. Ref.2
Phosphatase activity is slightly inhibited by ADP, NADPH and ATP, and moderately inhibited by NAD and 5'-AMP. Kinase activity is slightly inhibited by ADP and NADP. Ref.2
The phosphatase is inert toward the substrates of NAD kinase (NAD, NADH, ATP, and poly(P)). This demonstrates that the phosphatase activity never interferes with the NAD kinase activity by degrading its substrates (Ref.2).
In the N-terminal section; belongs to the inositol monophosphatase family.
In the C-terminal section; belongs to the NAD kinase family.
Kcat is 399 sec(-1) for kinase activity with ATP as substrate. Kcat is 424 sec(-1) for kinase activity with NAD as substrate. Kcat is 906 sec(-1) for phosphatase activity with fructose 1,6-bisphosphate and NADP as substrates. Kcat is 1007 sec(-1) for phosphatase activity with NADPH as substrate.
KM=0.35 mM for ATP (for kinase activity at pH 8.5 and at 85 degrees Celsius) Ref.2
KM=3 mM for NAD (for kinase activity at pH 8.5 and at 85 degrees Celsius)
Vmax=93.4 µmol/min/mg enzyme toward ATP (for kinase activity at pH 8.5 and at 85 degrees Celsius)
Vmax=99.2 µmol/min/mg enzyme toward NAD (for kinase activity at pH 8.5 and at 85 degrees Celsius)
Vmax=212 µmol/min/mg enzyme toward NADP (for phosphatase activity at pH 8.5 and at 85 degrees Celsius)
Vmax=212 µmol/min/mg enzyme toward fructose 1,6-bisphosphate (for phosphatase activity at pH 8.5 and at 85 degrees Celsius)
Vmax=236 µmol/min/mg enzyme toward NADPH (for phosphatase activity at pH 8.5 and at 85 degrees Celsius)
Optimum temperature is 100 degrees Celsius for phosphatase and kinase activies. Both are inactive below 60 degrees Celsius.
|Technical term||Complete proteome|
Direct protein sequencing
|Gene Ontology (GO)|
|Biological_process||NAD metabolic process|
Inferred from electronic annotation. Source: InterProNADP biosynthetic processdephosphorylationlysine metabolic process
Inferred from electronic annotation. Source: InterProphosphatidylinositol phosphorylation
Inferred from electronic annotation. Source: InterPro
Inferred from electronic annotation. Source: UniProtKB-SubCell
|Molecular_function||ATP bindingNAD bindingNAD+ kinase activityNADP phosphatase activitymetal ion binding|
Inferred from electronic annotation. Source: UniProtKB-KW
|Complete GO annotation...|
Sequence annotation (Features)
|Feature key||Position(s)||Length||Description||Graphical view||Feature identifier|
|Chain||1 – 574||574||Bifunctional NADP phosphatase/NAD kinase HAMAP-Rule MF_00361||PRO_0000120700|
|Nucleotide binding||362 – 363||2||NAD By similarity|
|Nucleotide binding||436 – 437||2||NAD By similarity|
|Nucleotide binding||477 – 482||6||NAD By similarity|
|Region||1 – 297||297||NADP phosphatase HAMAP-Rule MF_00361|
|Region||302 – 574||273||NAD kinase HAMAP-Rule MF_00361|
|Active site||362||1||Proton acceptor By similarity|
|Metal binding||69||1||Magnesium 1 By similarity|
|Metal binding||87||1||Magnesium 1 By similarity|
|Metal binding||87||1||Magnesium 2 By similarity|
|Metal binding||90||1||Magnesium 2 By similarity|
|Metal binding||243||1||Magnesium 2 By similarity|
|Binding site||90||1||Substrate By similarity|
|Binding site||367||1||NAD By similarity|
|Binding site||447||1||NAD By similarity|
|Binding site||464||1||NAD By similarity|
|Binding site||466||1||NAD By similarity|
|Binding site||474||1||NAD; via carbonyl oxygen By similarity|
|||"Complete genome sequence of the methanogenic archaeon, Methanococcus jannaschii."|
Bult C.J., White O., Olsen G.J., Zhou L., Fleischmann R.D., Sutton G.G., Blake J.A., FitzGerald L.M., Clayton R.A., Gocayne J.D., Kerlavage A.R., Dougherty B.A., Tomb J.-F., Adams M.D., Reich C.I., Overbeek R., Kirkness E.F., Weinstock K.G. Venter J.C.
Science 273:1058-1073(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440.
|||"MJ0917 in archaeon Methanococcus jannaschii is a novel NADP phosphatase/NAD kinase."|
Kawai S., Fukuda C., Mukai T., Murata K.
J. Biol. Chem. 280:39200-39207(2005) [PubMed] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE OF 1-8, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, SUBSTRATE SPECIFICITY, ENZYME REGULATION, SUBUNIT.
|L77117 Genomic DNA. Translation: AAB98922.1.|
|RefSeq||NP_247912.1. NC_000909.1. |
3D structure databases
Protein-protein interaction databases
Protocols and materials databases
Genome annotation databases
|EnsemblBacteria||AAB98922; AAB98922; MJ_0917. |
Family and domain databases
|Gene3D||22.214.171.124. 1 hit. |
126.96.36.19930. 1 hit.
|HAMAP||MF_00361. NAD_kinase. |
|InterPro||IPR017438. ATP-NAD_kinase_dom_1. |
|PANTHER||PTHR20275. PTHR20275. 1 hit. |
|Pfam||PF00459. Inositol_P. 2 hits. |
PF01513. NAD_kinase. 1 hit.
|PIRSF||PIRSF036641. Bifunctional_PpnK_predicted. 1 hit. |
|PRINTS||PR00377. IMPHPHTASES. |
|SUPFAM||SSF111331. SSF111331. 1 hit. |
|PROSITE||PS00629. IMP_1. 1 hit. |
|Accession||Primary (citable) accession number: Q58327|
|Entry status||Reviewed (UniProtKB/Swiss-Prot)|
|Annotation program||Prokaryotic Protein Annotation Program|