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Q8U2I1 (PNPH_PYRFU) Reviewed, UniProtKB/Swiss-Prot

Last modified May 14, 2014. Version 65. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
Probable 6-oxopurine nucleoside phosphorylase

EC=2.4.2.1
Alternative name(s):
Purine nucleoside phosphorylase
Short name=PNP
Short name=PfPNP
Gene names
Ordered Locus Names:PF0853
OrganismPyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1) [Reference proteome] [HAMAP]
Taxonomic identifier186497 [NCBI]
Taxonomic lineageArchaeaEuryarchaeotaThermococciThermococcalesThermococcaceaePyrococcus

Protein attributes

Sequence length265 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Purine nucleoside phosphorylase which is highly specific for 6-oxopurine nucleosides. Cleaves guanosine or inosine to respective bases and sugar-1-phosphate molecules. Involved in purine salvage. Ref.2 Ref.3

Catalytic activity

Purine nucleoside + phosphate = purine + alpha-D-ribose 1-phosphate. HAMAP-Rule MF_01963

Pathway

Purine metabolism; purine nucleoside salvage. HAMAP-Rule MF_01963

Subunit structure

Homohexamer. Dimer of a homotrimer. Ref.2

Miscellaneous

Although this enzyme belongs to the family of MTA phosphorylases based on sequence homology, it has been shown that conserved amino acid substitutions in the substrate binding pocket convert the substrate specificity of this enzyme from 6-aminopurines to 6-oxopurines. It seems that P.furiosus has developed a specific enzyme (PF0853) for the metabolism of 6-oxo-purines, next to the canonical MTA phosphorylase PF0016.

Sequence similarities

Belongs to the PNP/MTAP phosphorylase family. MTAP subfamily.

Biophysicochemical properties

Kinetic parameters:

KM=322 µM for inosine

KM=122 µM for guanosine

KM=1454 µM for adenosine

pH dependence:

Optimum pH is 7.4. Active from pH 5 to 10.

Temperature dependence:

Optimum temperature is 120 degrees Celsius. Thermostable up to 133 degrees Celsius.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Initiator methionine11Removed HAMAP-Rule MF_01963
Chain2 – 265264Probable 6-oxopurine nucleoside phosphorylase HAMAP-Rule MF_01963
PRO_0000415086

Regions

Region49 – 502Phosphate binding By similarity
Region82 – 832Phosphate binding By similarity
Region211 – 2133Substrate binding By similarity

Sites

Binding site101Phosphate By similarity
Binding site1871Substrate; via amide nitrogen By similarity
Binding site1881Phosphate By similarity
Site1691Important for substrate specificity By similarity
Site2231Important for substrate specificity By similarity

Amino acid modifications

Disulfide bond136 ↔ 202 Ref.2
Disulfide bond162 ↔ 190 Ref.2
Disulfide bond254 ↔ 256 Ref.2

Experimental info

Mutagenesis1691E → S: Decreases catalytic activity for insosine and allows the enzyme to phosphorolytically cleave MTA with a catalytic efficiency about 4-fold higher than for inosine; when associated with D-211, D-213 and A-223. Ref.3
Mutagenesis2111N → D: Decreases catalytic activity for insosine and allows the enzyme to phosphorolytically cleave MTA with a catalytic efficiency about 4-fold higher than for inosine; when associated with S-169, D-213 and A-223. Ref.3
Mutagenesis2131A → D: Decreases catalytic activity for insosine and allows the enzyme to phosphorolytically cleave MTA with a catalytic efficiency about 4-fold higher than for inosine; when associated with S-169, D-211 and A-223. Ref.3
Mutagenesis2231H → A: Decreases catalytic activity for insosine and allows the enzyme to phosphorolytically cleave MTA with a catalytic efficiency about 4-fold higher than for inosine; when associated with S-169, D-211, and D-213. Ref.3
Mutagenesis2541C → S: Fully active, but reduces thermodynamic and kinetic stability of the enzyme; when associated with S-256.
Mutagenesis2561C → S: Fully active, but reduces thermodynamic and kinetic stability of the enzyme; when associated with S-254.

Sequences

Sequence LengthMass (Da)Tools
Q8U2I1 [UniParc].

Last modified June 1, 2002. Version 1.
Checksum: 19BF1EBBB2FEDBB8

FASTA26529,208
        10         20         30         40         50         60 
MPRIAIVGGS GVYDFPAENK REETVKTPYG EVKITVGVVG DEEVAFLARH GKGHSIPPHK 

        70         80         90        100        110        120 
INYRANIWAL YELGVERIIA TSAVGSMNPE MKPGDFVILD QIIDFTVSRP RTFYDGEESP 

       130        140        150        160        170        180 
HERKFVAHVD FTEPYCPEIR KALITAARNL GLPYHPRGTY VCTEGPRFET AAEIRAYRIL 

       190        200        210        220        230        240 
GGDVVGMTQC PEAILARELE MCYATVAIVT NYAAGMSGKK LTHSEVVELM QKKSEDIVKL 

       250        260 
ILAAIPLIPK ERRCGCKDAL KGATG 

« Hide

References

« Hide 'large scale' references
[1]"Divergence of the hyperthermophilic archaea Pyrococcus furiosus and P. horikoshii inferred from complete genomic sequences."
Maeder D.L., Weiss R.B., Dunn D.M., Cherry J.L., Gonzalez J.M., DiRuggiero J., Robb F.T.
Genetics 152:1299-1305(1999) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: ATCC 43587 / DSM 3638 / JCM 8422 / Vc1.
[2]"Biochemical and structural characterization of mammalian-like purine nucleoside phosphorylase from the Archaeon Pyrococcus furiosus."
Cacciapuoti G., Gorassini S., Mazzeo M.F., Siciliano R.A., Carbone V., Zappia V., Porcelli M.
FEBS J. 274:2482-2495(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE OF N-TERMINUS, FUNCTION, SUBUNIT, DISULFIDE BONDS.
[3]"Unraveling the structural and functional differences between purine nucleoside phosphorylase and 5'-deoxy-5'-methylthioadenosine phosphorylase from the archaeon Pyrococcus furiosus."
Cacciapuoti G., Marabotti A., Fuccio F., Porcelli M.
Biochim. Biophys. Acta 1814:1358-1366(2011) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, MUTAGENESIS OF GLU-169; ASN-211; ALA-213 AND HIS-223.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
AE009950 Genomic DNA. Translation: AAL80977.1.
RefSeqNP_578582.1. NC_003413.1.

3D structure databases

ProteinModelPortalQ8U2I1.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

STRING186497.PF0853.

Proteomic databases

PRIDEQ8U2I1.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAL80977; AAL80977; PF0853.
GeneID1468711.
KEGGpfu:PF0853.

Phylogenomic databases

eggNOGCOG0005.
HOGENOMHOG000228987.
KOK00772.
OMACTPFGKP.

Enzyme and pathway databases

BRENDA2.4.2.1. 321.
UniPathwayUPA00606.

Family and domain databases

Gene3D3.40.50.1580. 1 hit.
HAMAPMF_01963. MTAP.
InterProIPR010044. MTAP.
IPR000845. Nucleoside_phosphorylase_d.
IPR001369. PNP/MTAP.
IPR018099. Purine_phosphorylase-2_CS.
[Graphical view]
PANTHERPTHR11904. PTHR11904. 1 hit.
PfamPF01048. PNP_UDP_1. 1 hit.
[Graphical view]
SUPFAMSSF53167. SSF53167. 1 hit.
TIGRFAMsTIGR01694. MTAP. 1 hit.
PROSITEPS01240. PNP_MTAP_2. 1 hit.
[Graphical view]
ProtoNetSearch...

Entry information

Entry namePNPH_PYRFU
AccessionPrimary (citable) accession number: Q8U2I1
Entry history
Integrated into UniProtKB/Swiss-Prot: January 25, 2012
Last sequence update: June 1, 2002
Last modified: May 14, 2014
This is version 65 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PATHWAY comments

Index of metabolic and biosynthesis pathways