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P83772 (CRNA_PSEPU) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 39. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (3) | 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:
Creatinine amidohydrolase

EC=3.5.2.10
Alternative name(s):
Creatininase
Gene names
Name:crnA
OrganismPseudomonas putida (Arthrobacter siderocapsulatus)
Taxonomic identifier303 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaPseudomonadalesPseudomonadaceaePseudomonas

Protein attributes

Sequence length260 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Cyclic amidohydrolase that catalyzes the reversible conversion of creatinine to creatine. Is also active toward glycocyamidine, though the reaction rate is very low, but it is completely inert toward hydantoin and its derivatives. Ref.2

Catalytic activity

Creatinine + H2O = creatine. Ref.2 Ref.5

Cofactor

Binds 2 Zn2+ ions per subunit. The Zn2+ in the metal 1 binding site can be replaced with Mn2+; however, the second zinc in metal binding site 2 is much more tightly bound and can not be replaced. The enzyme with one zinc and one manganese ion is more active than that with two zinc ions. Ref.3 Ref.4

Enzyme regulation

Is markedly inactivated in vitro by heavy metal ions, N-bromosuccinimide, ethoxyformic anhydride, and dye-sensitized photooxidation. Ref.2

Pathway

Amine and polyamine degradation; creatinine degradation.

Subunit structure

Homohexamer; trimer of dimers. Ref.3 Ref.4

Miscellaneous

The proposed catalytic mechanism involves two water molecules. The first molecule is a hydroxide ion that is bound as a bridge between the two metal ions and attacks the carbonyl carbon of the substrate. The second water molecule, that is bound to the carboxyl group of Glu-122 and to the metal 1, functions as a proton donor in catalysis.

Sequence similarities

Belongs to the creatininase superfamily.

Biophysicochemical properties

Kinetic parameters:

KM=26 mM for creatinine Ref.2 Ref.5

KM=130 mM for creatine

KM=200 mM for glycocyamidine

Vmax=390 µmol/min/mg enzyme for the forward reaction (creatine formation)

Vmax=1510 µmol/min/mg enzyme for the reverse reaction (creatinine formation)

Vmax=3.7 µmol/min/mg enzyme with glycocyamidine as substrate

pH dependence:

Optimum pH is 7-9 for the forward and reverse reactions.

Temperature dependence:

Retains 75% of the activity after incubation at 75 degrees Celsius for 30 minutes.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Initiator methionine11Removed Probable
Chain2 – 260259Creatinine amidohydrolase
PRO_0000406934

Regions

Region174 – 1785Substrate binding

Sites

Metal binding341Zinc or manganese 1
Metal binding361Zinc 2; via tele nitrogen
Metal binding451Zinc 2
Metal binding451Zinc or manganese 1
Metal binding1201Zinc or manganese 1; via pros nitrogen
Metal binding1831Zinc 2
Binding site781Substrate; via carbonyl oxygen
Binding site1211Substrate; via amide nitrogen
Site1221Coordinates a catalytic water molecule

Experimental info

Mutagenesis1211Y → A: 30-fold decrease in catalytic efficiency. Ref.5
Mutagenesis1221E → Q: 700-fold decrease in catalytic efficiency. No ion in metal binding site 1. Ref.5
Mutagenesis1541W → A: Loss of activity. Ref.5
Mutagenesis1541W → F: 340-fold decrease in catalytic efficiency. Ref.5
Mutagenesis1741W → A: Nearly no activity. Ref.5
Mutagenesis1741W → F: 2-fold decrease in catalytic efficiency. Ref.5
Mutagenesis1781H → A: Loss of activity.
Mutagenesis1831E → Q: Loss of activity. Ref.5

Secondary structure

.......................................... 260
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P83772 [UniParc].

Last modified March 15, 2004. Version 1.
Checksum: E530A7513F57A762

FASTA26028,569
        10         20         30         40         50         60 
MSKSVFVGEL TWKEYEARVA AGDCVLMLPV GALEQHGHHM CMNVDVLLPT AVCKRVAERI 

        70         80         90        100        110        120 
GALVMPGLQY GYKSQQKSGG GNHFPGTTSL DGATLTGTVQ DIIRELARHG ARRLVLMNGH 

       130        140        150        160        170        180 
YENSMFIVEG IDLALRELRY AGIQDFKVVV LSYWDFVKDP AVIQQLYPEG FLGWDIEHGG 

       190        200        210        220        230        240 
VFETSLMLAL YPDLVDLDRV VDHPPATFPP YDVFPVDPAR TPAPGTLSSA KTASREKGEL 

       250        260 
ILEVCVQGIA DAIREEFPPT 

« Hide

References

[1]"Cloning of the creatinine amidohydrolase gene from Pseudomonas sp. PS-7."
Yamamoto K., Oka M., Kikuchi T., Emi S.
Biosci. Biotechnol. Biochem. 59:1331-1332(1995) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-6.
Strain: PS-7.
[2]"Creatinine amidohydrolase (creatininase) from Pseudomonas putida. Purification and some properties."
Rikitake K., Oka I., Ando M., Yoshimoto T., Tsuru D.
J. Biochem. 86:1109-1117(1979) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION.
Strain: C-83.
[3]"Crystal structure of creatininase from Pseudomonas putida: a novel fold and a case of convergent evolution."
Beuth B., Niefind K., Schomburg D.
J. Mol. Biol. 332:287-301(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 2-260 IN COMPLEX WITH ZINC, COFACTOR, SUBUNIT.
[4]"Crystal structures of creatininase reveal the substrate binding site and provide an insight into the catalytic mechanism."
Yoshimoto T., Tanaka N., Kanada N., Inoue T., Nakajima Y., Haratake M., Nakamura K.T., Xu Y., Ito K.
J. Mol. Biol. 337:399-416(2004) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF APOENZYME AND COMPLEXES WITH CREATINE; ZINC AND MANGANESE, COFACTOR, SUBUNIT, CATALYTIC MECHANISM.
[5]"Substitution of Glu122 by glutamine revealed the function of the second water molecule as a proton donor in the binuclear metal enzyme creatininase."
Yamashita K., Nakajima Y., Matsushita H., Nishiya Y., Yamazawa R., Wu Y.F., Matsubara F., Oyama H., Ito K., Yoshimoto T.
J. Mol. Biol. 396:1081-1096(2010) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.78 ANGSTROMS) OF WILD-TYPE IN COMPLEX WITH 1-METHYLGUANIDINE INHIBITOR AND MUTANTS ALA-154; PHE-154; PHE-174 AND GLN-122 IN COMPLEX WITH CREATINE; ZINC AND MANGANESE, CATALYTIC ACTIVITY, KINETIC PARAMETERS, CATALYTIC MECHANISM, MUTAGENESIS OF TYR-121; GLU-122; TRP-154; TRP-174 AND GLU-183.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
D45424 Genomic DNA. Translation: BAA08265.1.
PIRT48846.

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1J2TX-ray1.80A/B/C/D/E/F1-260[»]
1J2UX-ray1.85A/B/C/D/E/F1-260[»]
1Q3KX-ray2.10A/B/C/D/E/F2-260[»]
1V7ZX-ray1.60A/B/C/D/E/F1-260[»]
3A6DX-ray1.90A/B/C/D/E/F1-260[»]
3A6EX-ray2.00A/B/C/D/E/F1-260[»]
3A6FX-ray1.78A/B/C/D/E/F1-260[»]
3A6GX-ray2.00A/B/C/D/E/F1-260[»]
3A6HX-ray2.00A/B/C/D/E/F1-260[»]
3A6JX-ray2.00A/B/C/D/E/F1-260[»]
3A6KX-ray2.20A/B/C/D/E/F1-260[»]
3A6LX-ray2.00A/B/C/D/E/F1-260[»]
ProteinModelPortalP83772.
SMRP83772. Positions 3-259.
ModBaseSearch...
MobiDBSearch...

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Enzyme and pathway databases

BioCycMetaCyc:MONOMER-10962.
UniPathwayUPA00274.

Family and domain databases

Gene3D3.40.50.10310. 1 hit.
InterProIPR024087. Creatininase-like_dom.
IPR003785. Creatininase/forma_Hydrolase.
[Graphical view]
PfamPF02633. Creatininase. 1 hit.
[Graphical view]
SUPFAMSSF102215. SSF102215. 1 hit.
ProtoNetSearch...

Other

EvolutionaryTraceP83772.

Entry information

Entry nameCRNA_PSEPU
AccessionPrimary (citable) accession number: P83772
Secondary accession number(s): Q52548
Entry history
Integrated into UniProtKB/Swiss-Prot: April 5, 2011
Last sequence update: March 15, 2004
Last modified: April 16, 2014
This is version 39 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

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

PATHWAY comments

Index of metabolic and biosynthesis pathways