PAP_GEOSE
ID PAP_GEOSE Reviewed; 148 AA.
AC S0F332;
DT 09-JUL-2014, integrated into UniProtKB/Swiss-Prot.
DT 18-SEP-2013, sequence version 1.
DT 03-AUG-2022, entry version 30.
DE RecName: Full=Protein-arginine-phosphatase;
DE Short=PAP;
DE EC=3.9.1.2 {ECO:0000269|PubMed:23770242};
DE AltName: Full=Phosphoarginine phosphatase;
GN Name=ywle;
OS Geobacillus stearothermophilus (Bacillus stearothermophilus).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Geobacillus.
OX NCBI_TaxID=1422;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 12980 / DSM 22 / CCM 2062 / JCM 2501 / NBRC 12550 / NCIMB 8923
RC / NCTC 10339 / R-35646 / VKM B-510;
RA Fuhrmann Jakob J.F., Clausen Tim T.C.;
RT "Characterization of the arginine phosphatase YwlE from Geobacillus
RT stearothermophilus.";
RL Submitted (JUN-2013) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP KINETIC PARAMETERS, ACTIVITY REGULATION, ACTIVE SITE, AND MUTAGENESIS OF
RP CYS-9 AND CYS-14.
RX PubMed=23838530; DOI=10.1021/cb4001469;
RA Fuhrmann J., Subramanian V., Thompson P.R.;
RT "Targeting the arginine phosphatase YwlE with a catalytic redox-based
RT inhibitor.";
RL ACS Chem. Biol. 8:2024-2032(2013).
RN [3]
RP FUNCTION AS AN ARGININE PHOSPHATASE, CATALYTIC ACTIVITY, SUBSTRATE
RP SPECIFICITY, KINETIC PARAMETERS, AND MUTAGENESIS OF THR-13.
RC STRAIN=ATCC 12980 / DSM 22 / CCM 2062 / JCM 2501 / NBRC 12550 / NCIMB 8923
RC / NCTC 10339 / R-35646 / VKM B-510;
RX PubMed=23770242; DOI=10.1016/j.celrep.2013.05.023;
RA Fuhrmann J., Mierzwa B., Trentini D.B., Spiess S., Lehner A.,
RA Charpentier E., Clausen T.;
RT "Structural basis for recognizing phosphoarginine and evolving residue-
RT specific protein phosphatases in gram-positive bacteria.";
RL Cell Rep. 3:1832-1839(2013).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) IN COMPLEX WITH PHOSPHATE,
RP BIOTECHNOLOGY, SUBUNIT, AND MUTAGENESIS OF CYS-9 AND ASP-117.
RX PubMed=24825175; DOI=10.1074/mcp.o113.035790;
RA Trentini D.B., Fuhrmann J., Mechtler K., Clausen T.;
RT "Chasing phosphoarginine proteins: development of a selective enrichment
RT method using a phosphatase trap.";
RL Mol. Cell. Proteomics 13:1953-1964(2014).
CC -!- FUNCTION: Catalyzes the specific dephosphorylation of phosphoarginine
CC residues in proteins. Probably counteracts the protein arginine kinase
CC McsB in vivo. Exhibits almost no activity against pTyr peptides.
CC Protein arginine phosphorylation has a physiologically important role
CC and is involved in the regulation of many critical cellular processes,
CC such as protein homeostasis, motility, competence, and stringent and
CC stress responses, by regulating gene expression and protein activity.
CC {ECO:0000269|PubMed:23770242}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N(omega)-phospho-L-arginyl-[protein] = L-arginyl-
CC [protein] + phosphate; Xref=Rhea:RHEA:43380, Rhea:RHEA-COMP:10532,
CC Rhea:RHEA-COMP:10533, ChEBI:CHEBI:15377, ChEBI:CHEBI:29965,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:83226; EC=3.9.1.2;
CC Evidence={ECO:0000269|PubMed:23770242};
CC -!- ACTIVITY REGULATION: Irreversibly inhibited by the synthetic inhibitor
CC cyc-SeCN-amidine, which inactivates the enzyme by inducing disulfide
CC bond formation between the two active site cysteine residues Cys-9 and
CC Cys-14. {ECO:0000269|PubMed:23838530}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=100 mM for p-nitrophenyl-phosphate (at 25 degrees Celsius and pH
CC 7.6) {ECO:0000269|PubMed:23770242, ECO:0000269|PubMed:23838530};
CC KM=124.52 mM for p-nitrophenyl-phosphate (at 40 degrees Celsius and
CC pH 8.0) {ECO:0000269|PubMed:23770242, ECO:0000269|PubMed:23838530};
CC Note=kcat is 1.12 sec(-1) with pNPP as substrate (at 25 degrees
CC Celsius and pH 7.6) (PubMed:23838530) and 3.2 sec(-1) with pNPP as
CC substrate (at 40 degrees Celsius and pH 8.0) (PubMed:23770242). pNPP
CC is a phospho-tyrosine mimicking compound.
CC {ECO:0000269|PubMed:23770242, ECO:0000269|PubMed:23838530};
CC -!- SUBUNIT: Is present in solution as a mixture of monomers, dimers and
CC higher order oligomers (trimers and tetramers).
CC {ECO:0000269|PubMed:24825175}.
CC -!- BIOTECHNOLOGY: An engineered YwlE Ala-9 mutant can be used as a
CC phosphatase trap that efficiently captures arginine-phosphorylated
CC proteins but cannot hydrolyze the captured substrates, thus
CC facilitating identification of phosphoarginine sites in the large pool
CC of cellular protein modifications. Therefore, this novel tool for the
CC selective enrichment and subsequent MS analysis of arginine
CC phosphorylation should facilitate the investigation of this highly
CC overlooked protein modification, in particular in eukaryotic samples.
CC {ECO:0000269|PubMed:24825175}.
CC -!- SIMILARITY: Belongs to the low molecular weight phosphotyrosine protein
CC phosphatase family. {ECO:0000305}.
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DR EMBL; HG316025; CDF77252.1; -; Genomic_DNA.
DR RefSeq; WP_033017317.1; NZ_RCTH01000008.1.
DR PDB; 4PIC; X-ray; 1.40 A; A/B=1-148.
DR PDBsum; 4PIC; -.
DR AlphaFoldDB; S0F332; -.
DR SMR; S0F332; -.
DR KEGG; ag:CDF77252; -.
DR BRENDA; 3.9.1.2; 623.
DR GO; GO:0098627; F:protein arginine phosphatase activity; IEA:UniProtKB-EC.
DR GO; GO:0004725; F:protein tyrosine phosphatase activity; IEA:InterPro.
DR GO; GO:0006470; P:protein dephosphorylation; IEA:InterPro.
DR InterPro; IPR023485; Ptyr_pPase.
DR InterPro; IPR036196; Ptyr_pPase_sf.
DR InterPro; IPR017867; Tyr_phospatase_low_mol_wt.
DR Pfam; PF01451; LMWPc; 1.
DR PRINTS; PR00719; LMWPTPASE.
DR SMART; SM00226; LMWPc; 1.
DR SUPFAM; SSF52788; SSF52788; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Protein phosphatase.
FT CHAIN 1..148
FT /note="Protein-arginine-phosphatase"
FT /id="PRO_0000429921"
FT ACT_SITE 9
FT /note="Nucleophile"
FT /evidence="ECO:0000269|PubMed:23838530"
FT ACT_SITE 15
FT /evidence="ECO:0000250|UniProtKB:P11064"
FT ACT_SITE 117
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:P11064"
FT BINDING 10..15
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT SITE 13
FT /note="Important for substrate discrimination"
FT MUTAGEN 9
FT /note="C->A: Mutant stably binds to the arginine-
FT phosphorylated substrate."
FT /evidence="ECO:0000269|PubMed:23838530,
FT ECO:0000269|PubMed:24825175"
FT MUTAGEN 9
FT /note="C->S: Complete loss of phosphatase activity."
FT /evidence="ECO:0000269|PubMed:23838530,
FT ECO:0000269|PubMed:24825175"
FT MUTAGEN 13
FT /note="T->I: Strongly decreases p-Arg phosphatase activity
FT and markedly increases p-Tyr phosphatase activity."
FT /evidence="ECO:0000269|PubMed:23770242"
FT MUTAGEN 14
FT /note="C->S: 6-fold reduction in phosphatase activity with
FT pNPP as substrate. Nearly no effect on substrate affinity."
FT /evidence="ECO:0000269|PubMed:23838530"
FT MUTAGEN 117
FT /note="D->A,N: Mutant does not bind to the arginine-
FT phosphorylated substrate."
FT /evidence="ECO:0000269|PubMed:24825175"
FT STRAND 3..14
FT /evidence="ECO:0007829|PDB:4PIC"
FT HELIX 15..25
FT /evidence="ECO:0007829|PDB:4PIC"
FT STRAND 31..37
FT /evidence="ECO:0007829|PDB:4PIC"
FT HELIX 48..56
FT /evidence="ECO:0007829|PDB:4PIC"
FT HELIX 70..75
FT /evidence="ECO:0007829|PDB:4PIC"
FT STRAND 77..83
FT /evidence="ECO:0007829|PDB:4PIC"
FT HELIX 84..93
FT /evidence="ECO:0007829|PDB:4PIC"
FT HELIX 95..100
FT /evidence="ECO:0007829|PDB:4PIC"
FT STRAND 101..103
FT /evidence="ECO:0007829|PDB:4PIC"
FT HELIX 104..109
FT /evidence="ECO:0007829|PDB:4PIC"
FT HELIX 123..148
FT /evidence="ECO:0007829|PDB:4PIC"
SQ SEQUENCE 148 AA; 16300 MW; 7C321C8E3F888D14 CRC64;
MPYRILFVCT GNTCRSPMAA ALLENKQLPG VEVKSAGVFA AEGSEASVHA KMVLKEKGIE
AAHRSSQLKK EHIDWATHVL AMTSGHKDMI VERFPEAKDK TFTLKQFVSG TDGDIADPFG
GPIEVYRAAR DELETLIDRL AEKLQTEQ
