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3a6j

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Revision as of 22:32, 25 December 2014

E122Q mutant creatininase complexed with creatine

Structural highlights

3a6j is a 6 chain structure with sequence from "bacillus_fluorescens_putidus"_flugge_1886 "bacillus fluorescens putidus" flugge 1886. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:	, ,
Related:	1j2t, 1j2u, 1v7z, 3a6d, 3a6e, 3a6f, 3a6g, 3a6h, 3a6k, 3a6l
Activity:	Creatininase, with EC number 3.5.2.10
Resources:	FirstGlance, OCA, RCSB, PDBsum

Function

[CRNA_PSEPU] 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.^[1]

Evolutionary Conservation

Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.

Publication Abstract from PubMed

Creatininase is a binuclear zinc enzyme and catalyzes the reversible conversion of creatinine to creatine. It exhibits an open-closed conformational change upon substrate binding, and the differences in the conformations of Tyr121, Trp154, and the loop region containing Trp174 were evident in the enzyme-creatine complex when compared to those in the ligand-free enzyme. We have determined the crystal structure of the enzyme complexed with a 1-methylguanidine. All subunits in the complex existed as the closed form, and the binding mode of creatinine was estimated. Site-directed mutagenesis revealed that the hydrophobic residues that show conformational change upon substrate binding are important for the enzyme activity. We propose a catalytic mechanism of creatininase in which two water molecules have significant roles. The first molecule is a hydroxide ion (Wat1) that is bound as a bridge between the two metal ions and attacks the carbonyl carbon of the substrate. The second molecule is a water molecule (Wat2) that is bound to the carboxyl group of Glu122 and functions as a proton donor in catalysis. The activity of the E122Q mutant was very low and it was only partially restored by the addition of ZnCl(2) or MnCl(2). In the E122Q mutant, k(cat) is drastically decreased, indicating that Glu122 is important for catalysis. X-ray crystallographic study and the atomic absorption spectrometry analysis of the E122Q mutant-substrate complex revealed that the drastic decrease of the activity of the E122Q was caused by not only the loss of one Zn ion at the Metal1 site but also a critical function of Glu122, which most likely exists for a proton transfer step through Wat2.

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 YF, Matsubara F, Oyama H, Ito K, Yoshimoto T J Mol Biol. 2010 Mar 5;396(4):1081-96. Epub 2010 Jan 4. PMID:20043918^[2]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Rikitake K, Oka I, Ando M, Yoshimoto T, Tsuru D. Creatinine amidohydrolase (creatininase) from Pseudomonas putida. Purification and some properties. J Biochem. 1979 Oct;86(4):1109-17. PMID:500580
↑ Yamashita K, Nakajima Y, Matsushita H, Nishiya Y, Yamazawa R, Wu YF, Matsubara F, Oyama H, Ito K, Yoshimoto T. Substitution of Glu122 by glutamine revealed the function of the second water molecule as a proton donor in the binuclear metal enzyme creatininase. J Mol Biol. 2010 Mar 5;396(4):1081-96. Epub 2010 Jan 4. PMID:20043918 doi:10.1016/j.jmb.2009.12.045

1 Structural highlights
2 Function
3 Evolutionary Conservation
4 Publication Abstract from PubMed
5 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/3a6j"

@@ Line 8: / Line 8: @@
 <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3a6j FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3a6j OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3a6j RCSB], [http://www.ebi.ac.uk/pdbsum/3a6j PDBsum]</span></td></tr>
 </table>
+== Function ==
+[[http://www.uniprot.org/uniprot/CRNA_PSEPU CRNA_PSEPU]] 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>PMID:500580</ref>
 == Evolutionary Conservation ==
 [[Image:Consurf_key_small.gif|200px|right]]
@@ Line 32: / Line 34: @@
 [[Category: Bacillus fluorescens putidus flugge 1886]]
 [[Category: Creatininase]]
-[[Category: Ito, K.]]
+[[Category: Ito, K]]
-[[Category: Nakajima, Y.]]
+[[Category: Nakajima, Y]]
-[[Category: Yamashita, K.]]
+[[Category: Yamashita, K]]
-[[Category: Yoshimoto, T.]]
+[[Category: Yoshimoto, T]]
 [[Category: Creatinine amidohydrolase]]
 [[Category: Hydrolase]]
 [[Category: Urease-related amidohydrolase superfamily]]

3a6j

From Proteopedia

Revision as of 22:32, 25 December 2014

E122Q mutant creatininase complexed with creatine

Structural highlights

Function

Evolutionary Conservation

Publication Abstract from PubMed

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

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