CRNA_PSEPU
ID CRNA_PSEPU Reviewed; 260 AA.
AC P83772; Q52548;
DT 05-APR-2011, integrated into UniProtKB/Swiss-Prot.
DT 15-MAR-2004, sequence version 1.
DT 03-AUG-2022, entry version 65.
DE RecName: Full=Creatinine amidohydrolase;
DE EC=3.5.2.10;
DE AltName: Full=Creatininase;
GN Name=crnA;
OS Pseudomonas putida (Arthrobacter siderocapsulatus).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=303;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 2-6.
RC STRAIN=PS-7;
RX PubMed=7670196; DOI=10.1271/bbb.59.1331;
RA Yamamoto K., Oka M., Kikuchi T., Emi S.;
RT "Cloning of the creatinine amidohydrolase gene from Pseudomonas sp. PS-7.";
RL Biosci. Biotechnol. Biochem. 59:1331-1332(1995).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND ACTIVITY REGULATION.
RC STRAIN=C-83;
RX PubMed=500580; DOI=10.1093/oxfordjournals.jbchem.a132605;
RA Rikitake K., Oka I., Ando M., Yoshimoto T., Tsuru D.;
RT "Creatinine amidohydrolase (creatininase) from Pseudomonas putida.
RT Purification and some properties.";
RL J. Biochem. 86:1109-1117(1979).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 2-260 IN COMPLEX WITH ZINC,
RP COFACTOR, AND SUBUNIT.
RX PubMed=12946365; DOI=10.1016/s0022-2836(03)00860-x;
RA Beuth B., Niefind K., Schomburg D.;
RT "Crystal structure of creatininase from Pseudomonas putida: a novel fold
RT and a case of convergent evolution.";
RL J. Mol. Biol. 332:287-301(2003).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF APOENZYME AND COMPLEXES WITH
RP CREATINE; ZINC AND MANGANESE, COFACTOR, SUBUNIT, AND CATALYTIC MECHANISM.
RX PubMed=15003455; DOI=10.1016/j.jmb.2004.01.022;
RA Yoshimoto T., Tanaka N., Kanada N., Inoue T., Nakajima Y., Haratake M.,
RA Nakamura K.T., Xu Y., Ito K.;
RT "Crystal structures of creatininase reveal the substrate binding site and
RT provide an insight into the catalytic mechanism.";
RL J. Mol. Biol. 337:399-416(2004).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.78 ANGSTROMS) OF WILD-TYPE IN COMPLEX WITH
RP 1-METHYLGUANIDINE INHIBITOR AND MUTANTS ALA-154; PHE-154; PHE-174 AND
RP GLN-122 IN COMPLEX WITH CREATINE; ZINC AND MANGANESE, CATALYTIC ACTIVITY,
RP KINETIC PARAMETERS, CATALYTIC MECHANISM, AND MUTAGENESIS OF TYR-121;
RP GLU-122; TRP-154; TRP-174 AND GLU-183.
RX PubMed=20043918; DOI=10.1016/j.jmb.2009.12.045;
RA Yamashita K., Nakajima Y., Matsushita H., Nishiya Y., Yamazawa R., Wu Y.F.,
RA Matsubara F., Oyama H., Ito K., Yoshimoto T.;
RT "Substitution of Glu122 by glutamine revealed the function of the second
RT water molecule as a proton donor in the binuclear metal enzyme
RT creatininase.";
RL J. Mol. Biol. 396:1081-1096(2010).
CC -!- FUNCTION: Cyclic amidohydrolase that catalyzes the reversible
CC conversion of creatinine to creatine. Is also active toward
CC glycocyamidine, though the reaction rate is very low, but it is
CC completely inert toward hydantoin and its derivatives.
CC {ECO:0000269|PubMed:500580}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=creatinine + H2O = creatine; Xref=Rhea:RHEA:14533,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:16737, ChEBI:CHEBI:57947; EC=3.5.2.10;
CC Evidence={ECO:0000269|PubMed:20043918, ECO:0000269|PubMed:500580};
CC -!- COFACTOR:
CC Note=Binds 2 Zn(2+) ions per subunit. The Zn(2+) in the metal 1 binding
CC site can be replaced with Mn(2+); however, the second zinc in metal
CC binding site 2 is much more tightly bound and cannot be replaced. The
CC enzyme with one zinc and one manganese ion is more active than that
CC with two zinc ions. {ECO:0000269|PubMed:12946365,
CC ECO:0000269|PubMed:15003455};
CC -!- ACTIVITY REGULATION: Is markedly inactivated in vitro by heavy metal
CC ions, N-bromosuccinimide, ethoxyformic anhydride, and dye-sensitized
CC photooxidation. {ECO:0000269|PubMed:500580}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=26 mM for creatinine {ECO:0000269|PubMed:20043918,
CC ECO:0000269|PubMed:500580};
CC KM=130 mM for creatine {ECO:0000269|PubMed:20043918,
CC ECO:0000269|PubMed:500580};
CC KM=200 mM for glycocyamidine {ECO:0000269|PubMed:20043918,
CC ECO:0000269|PubMed:500580};
CC Vmax=390 umol/min/mg enzyme for the forward reaction (creatine
CC formation) {ECO:0000269|PubMed:20043918, ECO:0000269|PubMed:500580};
CC Vmax=1510 umol/min/mg enzyme for the reverse reaction (creatinine
CC formation) {ECO:0000269|PubMed:20043918, ECO:0000269|PubMed:500580};
CC Vmax=3.7 umol/min/mg enzyme with glycocyamidine as substrate
CC {ECO:0000269|PubMed:20043918, ECO:0000269|PubMed:500580};
CC pH dependence:
CC Optimum pH is 7-9 for the forward and reverse reactions.
CC {ECO:0000269|PubMed:500580};
CC Temperature dependence:
CC Retains 75% of the activity after incubation at 75 degrees Celsius
CC for 30 minutes. {ECO:0000269|PubMed:500580};
CC -!- PATHWAY: Amine and polyamine degradation; creatinine degradation.
CC -!- SUBUNIT: Homohexamer; trimer of dimers. {ECO:0000269|PubMed:12946365,
CC ECO:0000269|PubMed:15003455, ECO:0000269|PubMed:20043918}.
CC -!- MISCELLANEOUS: The proposed catalytic mechanism involves two water
CC molecules. The first molecule is a hydroxide ion that is bound as a
CC bridge between the two metal ions and attacks the carbonyl carbon of
CC the substrate. The second water molecule, that is bound to the carboxyl
CC group of Glu-122 and to the metal 1, functions as a proton donor in
CC catalysis.
CC -!- SIMILARITY: Belongs to the creatininase superfamily. {ECO:0000305}.
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DR EMBL; D45424; BAA08265.1; -; Genomic_DNA.
DR PIR; T48846; T48846.
DR PDB; 1J2T; X-ray; 1.80 A; A/B/C/D/E/F=1-260.
DR PDB; 1J2U; X-ray; 1.85 A; A/B/C/D/E/F=1-260.
DR PDB; 1Q3K; X-ray; 2.10 A; A/B/C/D/E/F=2-260.
DR PDB; 1V7Z; X-ray; 1.60 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6D; X-ray; 1.90 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6E; X-ray; 2.00 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6F; X-ray; 1.78 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6G; X-ray; 2.00 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6H; X-ray; 2.00 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6J; X-ray; 2.00 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6K; X-ray; 2.20 A; A/B/C/D/E/F=1-260.
DR PDB; 3A6L; X-ray; 2.00 A; A/B/C/D/E/F=1-260.
DR PDBsum; 1J2T; -.
DR PDBsum; 1J2U; -.
DR PDBsum; 1Q3K; -.
DR PDBsum; 1V7Z; -.
DR PDBsum; 3A6D; -.
DR PDBsum; 3A6E; -.
DR PDBsum; 3A6F; -.
DR PDBsum; 3A6G; -.
DR PDBsum; 3A6H; -.
DR PDBsum; 3A6J; -.
DR PDBsum; 3A6K; -.
DR PDBsum; 3A6L; -.
DR AlphaFoldDB; P83772; -.
DR SMR; P83772; -.
DR BioCyc; MetaCyc:MON-10962; -.
DR BRENDA; 3.5.2.10; 5092.
DR SABIO-RK; P83772; -.
DR UniPathway; UPA00274; -.
DR EvolutionaryTrace; P83772; -.
DR GO; GO:0047789; F:creatininase activity; IDA:UniProtKB.
DR GO; GO:0030145; F:manganese ion binding; IDA:UniProtKB.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
DR GO; GO:0006601; P:creatine biosynthetic process; IDA:UniProtKB.
DR GO; GO:0006602; P:creatinine catabolic process; IDA:UniProtKB.
DR Gene3D; 3.40.50.10310; -; 1.
DR InterPro; IPR031034; Creatininase.
DR InterPro; IPR024087; Creatininase-like_sf.
DR InterPro; IPR003785; Creatininase/forma_Hydrolase.
DR PANTHER; PTHR35005; PTHR35005; 1.
DR Pfam; PF02633; Creatininase; 1.
DR SUPFAM; SSF102215; SSF102215; 1.
DR TIGRFAMs; TIGR04448; creatininase; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Hydrolase; Manganese;
KW Metal-binding; Zinc.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000305|PubMed:7670196"
FT CHAIN 2..260
FT /note="Creatinine amidohydrolase"
FT /id="PRO_0000406934"
FT BINDING 34
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:20043918"
FT BINDING 34
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12946365,
FT ECO:0000269|PubMed:20043918"
FT BINDING 36
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12946365,
FT ECO:0000269|PubMed:20043918"
FT BINDING 45
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:20043918"
FT BINDING 45
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12946365,
FT ECO:0000269|PubMed:20043918"
FT BINDING 45
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12946365,
FT ECO:0000269|PubMed:20043918"
FT BINDING 78
FT /ligand="substrate"
FT BINDING 120
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:20043918"
FT BINDING 120
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12946365,
FT ECO:0000269|PubMed:20043918"
FT BINDING 121
FT /ligand="substrate"
FT BINDING 174..178
FT /ligand="substrate"
FT BINDING 183
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12946365,
FT ECO:0000269|PubMed:20043918"
FT SITE 122
FT /note="Coordinates a catalytic water molecule"
FT MUTAGEN 121
FT /note="Y->A: 30-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:20043918"
FT MUTAGEN 122
FT /note="E->Q: 700-fold decrease in catalytic efficiency. No
FT ion in metal binding site 1."
FT /evidence="ECO:0000269|PubMed:20043918"
FT MUTAGEN 154
FT /note="W->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:20043918"
FT MUTAGEN 154
FT /note="W->F: 340-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:20043918"
FT MUTAGEN 174
FT /note="W->A: Nearly no activity."
FT /evidence="ECO:0000269|PubMed:20043918"
FT MUTAGEN 174
FT /note="W->F: 2-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:20043918"
FT MUTAGEN 178
FT /note="H->A: Loss of activity."
FT MUTAGEN 183
FT /note="E->Q: Loss of activity."
FT /evidence="ECO:0000269|PubMed:20043918"
FT HELIX 7..9
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 12..20
FT /evidence="ECO:0007829|PDB:1V7Z"
FT STRAND 26..30
FT /evidence="ECO:0007829|PDB:1V7Z"
FT STRAND 38..40
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 44..60
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 76..79
FT /evidence="ECO:0007829|PDB:1V7Z"
FT STRAND 84..86
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 92..109
FT /evidence="ECO:0007829|PDB:1V7Z"
FT STRAND 113..118
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 121..123
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 124..140
FT /evidence="ECO:0007829|PDB:1V7Z"
FT STRAND 147..152
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 153..156
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 160..166
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 174..176
FT /evidence="ECO:0007829|PDB:3A6F"
FT STRAND 178..180
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 181..190
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 192..194
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 197..199
FT /evidence="ECO:0007829|PDB:1V7Z"
FT STRAND 210..215
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 218..220
FT /evidence="ECO:0007829|PDB:1V7Z"
FT HELIX 235..256
FT /evidence="ECO:0007829|PDB:1V7Z"
SQ SEQUENCE 260 AA; 28569 MW; E530A7513F57A762 CRC64;
MSKSVFVGEL TWKEYEARVA AGDCVLMLPV GALEQHGHHM CMNVDVLLPT AVCKRVAERI
GALVMPGLQY GYKSQQKSGG GNHFPGTTSL DGATLTGTVQ DIIRELARHG ARRLVLMNGH
YENSMFIVEG IDLALRELRY AGIQDFKVVV LSYWDFVKDP AVIQQLYPEG FLGWDIEHGG
VFETSLMLAL YPDLVDLDRV VDHPPATFPP YDVFPVDPAR TPAPGTLSSA KTASREKGEL
ILEVCVQGIA DAIREEFPPT
