PNPH_PYRFU
ID PNPH_PYRFU Reviewed; 265 AA.
AC Q8U2I1;
DT 25-JAN-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2002, sequence version 1.
DT 03-AUG-2022, entry version 100.
DE RecName: Full=Probable 6-oxopurine nucleoside phosphorylase {ECO:0000255|HAMAP-Rule:MF_01963};
DE EC=2.4.2.1 {ECO:0000255|HAMAP-Rule:MF_01963};
DE AltName: Full=Purine nucleoside phosphorylase {ECO:0000255|HAMAP-Rule:MF_01963};
DE Short=PNP {ECO:0000255|HAMAP-Rule:MF_01963};
DE Short=PfPNP;
GN OrderedLocusNames=PF0853;
OS Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1).
OC Archaea; Euryarchaeota; Thermococci; Thermococcales; Thermococcaceae;
OC Pyrococcus.
OX NCBI_TaxID=186497;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 43587 / DSM 3638 / JCM 8422 / Vc1;
RX PubMed=10430560; DOI=10.1093/genetics/152.4.1299;
RA Maeder D.L., Weiss R.B., Dunn D.M., Cherry J.L., Gonzalez J.M.,
RA DiRuggiero J., Robb F.T.;
RT "Divergence of the hyperthermophilic archaea Pyrococcus furiosus and P.
RT horikoshii inferred from complete genomic sequences.";
RL Genetics 152:1299-1305(1999).
RN [2]
RP PROTEIN SEQUENCE OF N-TERMINUS, FUNCTION, SUBUNIT, AND DISULFIDE BONDS.
RX PubMed=17419725; DOI=10.1111/j.1742-4658.2007.05784.x;
RA Cacciapuoti G., Gorassini S., Mazzeo M.F., Siciliano R.A., Carbone V.,
RA Zappia V., Porcelli M.;
RT "Biochemical and structural characterization of mammalian-like purine
RT nucleoside phosphorylase from the Archaeon Pyrococcus furiosus.";
RL FEBS J. 274:2482-2495(2007).
RN [3]
RP FUNCTION, AND MUTAGENESIS OF GLU-169; ASN-211; ALA-213 AND HIS-223.
RX PubMed=21683167; DOI=10.1016/j.bbapap.2011.06.001;
RA Cacciapuoti G., Marabotti A., Fuccio F., Porcelli M.;
RT "Unraveling the structural and functional differences between purine
RT nucleoside phosphorylase and 5'-deoxy-5'-methylthioadenosine phosphorylase
RT from the archaeon Pyrococcus furiosus.";
RL Biochim. Biophys. Acta 1814:1358-1366(2011).
CC -!- FUNCTION: Purine nucleoside phosphorylase which is highly specific for
CC 6-oxopurine nucleosides. Cleaves guanosine or inosine to respective
CC bases and sugar-1-phosphate molecules. Involved in purine salvage.
CC {ECO:0000255|HAMAP-Rule:MF_01963, ECO:0000269|PubMed:17419725,
CC ECO:0000269|PubMed:21683167}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a purine D-ribonucleoside + phosphate = a purine nucleobase +
CC alpha-D-ribose 1-phosphate; Xref=Rhea:RHEA:19805, ChEBI:CHEBI:26386,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:57720, ChEBI:CHEBI:142355; EC=2.4.2.1;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01963};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=322 uM for inosine;
CC KM=122 uM for guanosine;
CC KM=1454 uM for adenosine;
CC pH dependence:
CC Optimum pH is 7.4. Active from pH 5 to 10.;
CC Temperature dependence:
CC Optimum temperature is 120 degrees Celsius. Thermostable up to 133
CC degrees Celsius.;
CC -!- PATHWAY: Purine metabolism; purine nucleoside salvage.
CC {ECO:0000255|HAMAP-Rule:MF_01963}.
CC -!- SUBUNIT: Homohexamer. Dimer of a homotrimer. {ECO:0000255|HAMAP-
CC Rule:MF_01963, ECO:0000269|PubMed:17419725}.
CC -!- MISCELLANEOUS: Although this enzyme belongs to the family of MTA
CC phosphorylases based on sequence homology, it has been shown that
CC conserved amino acid substitutions in the substrate binding pocket
CC convert the substrate specificity of this enzyme from 6-aminopurines to
CC 6-oxopurines. It seems that P.furiosus has developed a specific enzyme
CC (PF0853) for the metabolism of 6-oxo-purines, next to the canonical MTA
CC phosphorylase PF0016.
CC -!- SIMILARITY: Belongs to the PNP/MTAP phosphorylase family. MTAP
CC subfamily. {ECO:0000255|HAMAP-Rule:MF_01963}.
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DR EMBL; AE009950; AAL80977.1; -; Genomic_DNA.
DR RefSeq; WP_011011984.1; NZ_CP023154.1.
DR AlphaFoldDB; Q8U2I1; -.
DR SMR; Q8U2I1; -.
DR STRING; 186497.PF0853; -.
DR EnsemblBacteria; AAL80977; AAL80977; PF0853.
DR GeneID; 41712658; -.
DR KEGG; pfu:PF0853; -.
DR PATRIC; fig|186497.12.peg.903; -.
DR eggNOG; arCOG01327; Archaea.
DR HOGENOM; CLU_054456_0_2_2; -.
DR OMA; MTQCPEA; -.
DR OrthoDB; 63298at2157; -.
DR PhylomeDB; Q8U2I1; -.
DR BRENDA; 2.4.2.1; 5243.
DR UniPathway; UPA00606; -.
DR Proteomes; UP000001013; Chromosome.
DR GO; GO:0017061; F:S-methyl-5-thioadenosine phosphorylase activity; IEA:InterPro.
DR GO; GO:0006166; P:purine ribonucleoside salvage; IEA:UniProtKB-UniRule.
DR CDD; cd09010; MTAP_SsMTAPII_like_MTIP; 1.
DR Gene3D; 3.40.50.1580; -; 1.
DR HAMAP; MF_01963; MTAP; 1.
DR InterPro; IPR010044; MTAP.
DR InterPro; IPR000845; Nucleoside_phosphorylase_d.
DR InterPro; IPR035994; Nucleoside_phosphorylase_sf.
DR InterPro; IPR018099; Purine_phosphorylase-2_CS.
DR PANTHER; PTHR42679; PTHR42679; 1.
DR Pfam; PF01048; PNP_UDP_1; 1.
DR SUPFAM; SSF53167; SSF53167; 1.
DR TIGRFAMs; TIGR01694; MTAP; 1.
DR PROSITE; PS01240; PNP_MTAP_2; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Disulfide bond; Glycosyltransferase;
KW Purine salvage; Reference proteome; Transferase.
FT INIT_MET 1
FT /note="Removed"
FT CHAIN 2..265
FT /note="Probable 6-oxopurine nucleoside phosphorylase"
FT /id="PRO_0000415086"
FT BINDING 10
FT /ligand="phosphate"
FT /ligand_id="ChEBI:CHEBI:43474"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT BINDING 49..50
FT /ligand="phosphate"
FT /ligand_id="ChEBI:CHEBI:43474"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT BINDING 82..83
FT /ligand="phosphate"
FT /ligand_id="ChEBI:CHEBI:43474"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT BINDING 187
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT BINDING 188
FT /ligand="phosphate"
FT /ligand_id="ChEBI:CHEBI:43474"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT BINDING 211..213
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT SITE 169
FT /note="Important for substrate specificity"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT SITE 223
FT /note="Important for substrate specificity"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01963"
FT DISULFID 136..202
FT /evidence="ECO:0000269|PubMed:17419725"
FT DISULFID 162..190
FT /evidence="ECO:0000269|PubMed:17419725"
FT DISULFID 254..256
FT /evidence="ECO:0000269|PubMed:17419725"
FT MUTAGEN 169
FT /note="E->S: Decreases catalytic activity for insosine and
FT allows the enzyme to phosphorolytically cleave MTA with a
FT catalytic efficiency about 4-fold higher than for inosine;
FT when associated with D-211, D-213 and A-223."
FT /evidence="ECO:0000269|PubMed:21683167"
FT MUTAGEN 211
FT /note="N->D: Decreases catalytic activity for insosine and
FT allows the enzyme to phosphorolytically cleave MTA with a
FT catalytic efficiency about 4-fold higher than for inosine;
FT when associated with S-169, D-213 and A-223."
FT /evidence="ECO:0000269|PubMed:21683167"
FT MUTAGEN 213
FT /note="A->D: Decreases catalytic activity for insosine and
FT allows the enzyme to phosphorolytically cleave MTA with a
FT catalytic efficiency about 4-fold higher than for inosine;
FT when associated with S-169, D-211 and A-223."
FT /evidence="ECO:0000269|PubMed:21683167"
FT MUTAGEN 223
FT /note="H->A: Decreases catalytic activity for insosine and
FT allows the enzyme to phosphorolytically cleave MTA with a
FT catalytic efficiency about 4-fold higher than for inosine;
FT when associated with S-169, D-211, and D-213."
FT /evidence="ECO:0000269|PubMed:21683167"
FT MUTAGEN 254
FT /note="C->S: Fully active, but reduces thermodynamic and
FT kinetic stability of the enzyme; when associated with S-
FT 256."
FT MUTAGEN 256
FT /note="C->S: Fully active, but reduces thermodynamic and
FT kinetic stability of the enzyme; when associated with S-
FT 254."
SQ SEQUENCE 265 AA; 29208 MW; 19BF1EBBB2FEDBB8 CRC64;
MPRIAIVGGS GVYDFPAENK REETVKTPYG EVKITVGVVG DEEVAFLARH GKGHSIPPHK
INYRANIWAL YELGVERIIA TSAVGSMNPE MKPGDFVILD QIIDFTVSRP RTFYDGEESP
HERKFVAHVD FTEPYCPEIR KALITAARNL GLPYHPRGTY VCTEGPRFET AAEIRAYRIL
GGDVVGMTQC PEAILARELE MCYATVAIVT NYAAGMSGKK LTHSEVVELM QKKSEDIVKL
ILAAIPLIPK ERRCGCKDAL KGATG
