DB code: D00244

RLCP classification 4.12.18410.92 : Addition
5.201.672500.74 : Elimination
CATH domain 3.40.350.10 : Creatine Amidinohydrolase; Chain A, domain 1
3.90.230.10 : Creatine Amidinohydrolase Catalytic domain
E.C. 3.5.3.3
CSA 1chm
M-CSA 1chm
MACiE M0096

CATH domain Related DB codes (homologues)
3.40.350.10 : Creatine Amidinohydrolase; Chain A, domain 1 D00185
3.90.230.10 : Creatine Amidinohydrolase D00185

Uniprot Enzyme Name
UniprotKB Protein name Synonyms Pfam
P38488 Creatinase
EC 3.5.3.3
Creatine amidinohydrolase
PF01321 (Creatinase_N)
PF00557 (Peptidase_M24)
[Graphical View]

KEGG enzyme name
creatinase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P38488 CREA_PSEPU Creatine + H(2)O = sarcosine + urea. Homodimer.

KEGG Pathways
Map code Pathways E.C.
MAP00330 Arginine and proline metabolism

Compound table
Substrates Products Intermediates
KEGG-id C00300 C00001 C00213 C00086
E.C.
Compound Creatine H2O Sarcosine Urea Tetrahedral intermediate
Type amino acids,amide group,imine group H2O amino acids amide group,amine group
ChEBI 16919
 57947
 16919
 57947
 		15377
 15377
 		15611
 57433
 15611
 57433
 		16199
 48376
 16199
 48376
PubChem 586
 59098743
 586
 59098743
 		22247451
 962
 22247451
 962
 		1088
 7311726
 1088
 7311726
 		1176
 1176
1chmA01 Unbound Unbound Unbound Unbound
1chmB01 Unbound Unbound Unbound Unbound
1chmA02 Unbound Unbound Unbound Intermediate-analogue:CMS
1chmB02 Analogue:CMS Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
literature [1], [2]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1chmA01
1chmB01
1chmA02 HIS 232;GLU 262;GLU 358
1chmB02 HIS 232;GLU 262;GLU 358

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[1]
Fig.11, Fig.12, p.430-432
[2]
Figure 10, p.607 4

References
[1]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS).
Medline ID 89125596
PubMed ID 3221393
Journal J Mol Biol
Year 1988
Volume 204
Pages 417-33
Authors Hoeffken HW, Knof SH, Bartlett PA, Huber R, Moellering H, Schumacher G
Title Crystal structure determination, refinement and molecular model of creatine amidinohydrolase from Pseudomonas putida.
Related PDB
Related UniProtKB P38488
[2]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS).
Medline ID 90339496
PubMed ID 1696320
Journal J Mol Biol
Year 1990
Volume 214
Pages 597-610
Authors Coll M, Knof SH, Ohga Y, Messerschmidt A, Huber R, Moellering H, Russmann L, Schumacher G
Title Enzymatic mechanism of creatine amidinohydrolase as deduced from crystal structures.
Related PDB 1chm
Related UniProtKB P38488
[3]
Resource
Comments
Medline ID
PubMed ID 8504814
Journal Eur J Biochem
Year 1993
Volume 213
Pages 1225-33
Authors Schumann J, Jaenicke R
Title Creatinase in its collapsed A state shows properties of a molten globule with dimeric quaternary structure.
Related PDB
Related UniProtKB
[4]
Resource
Comments
Medline ID
PubMed ID 8251936
Journal Protein Sci
Year 1993
Volume 2
Pages 1612-20
Authors Schumann J, Bohm G, Schumacher G, Rudolph R, Jaenicke R
Title Stabilization of creatinase from Pseudomonas putida by random mutagenesis.
Related PDB
Related UniProtKB
[5]
Resource
Comments
Medline ID
PubMed ID 8291080
Journal Trends Biochem Sci
Year 1993
Volume 18
Pages 403-5
Authors Murzin AG
Title Can homologous proteins evolve different enzymatic activities?
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 8146141
Journal Proc Natl Acad Sci U S A
Year 1994
Volume 91
Pages 2473-7
Authors Bazan JF, Weaver LH, Roderick SL, Huber R, Matthews BW
Title Sequence and structure comparison suggest that methionine aminopeptidase, prolidase, aminopeptidase P, and creatinase share a common fold.
Related PDB
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 10387007
Journal Biochemistry
Year 1999
Volume 38
Pages 7678-88
Authors Lowther WT, Orville AM, Madden DT, Lim S, Rich DH, Matthews BW
Title Escherichia coli methionine aminopeptidase: implications of crystallographic analyses of the native, mutant, and inhibited enzymes for the mechanism of catalysis.
Related PDB
Related UniProtKB

Comments
Although this enzyme belongs to the peptidase family-M24, it does not utilize a divalent metal ion as a cofactor.
According to the literature [2], this enzyme catalyzes two successive reactions, addition and elimination, as follows:
(A) Addition of water to double-bond:
(A0) Throughout this reaction, Glu262 and Glu358 stabilize the positive charge on the guanidinium group.
(A1) His232 acts as a general base to deprotonate and activate the water.
(A2) The activated water or hydroxyl group makes a nucleophilic attack on the C(1) carbon of guanidium group (addition site), leading to the formation of tetrahedral intermediate.
(A3) His232 acts as a general acid to protonate the N(3) nitrogen atom (protonation site), forming guanidinium hydrate.
(B) Eliminative double-bond formation:
(B1) His232 acts as a general base to deptoronate guanidinium hydrate (deprotonation site), leading to the cleavage of the covalent bond between C(1) and N(3) atoms.

Created Updated
2006-04-04 2009-02-26