1n31
From Proteopedia
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| - | + | {{STRUCTURE_1n31| PDB=1n31 | SCENE= }} | |
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'''Structure of A Catalytically Inactive Mutant (K223A) of C-DES with a Substrate (Cystine) Linked to the Co-Factor''' | '''Structure of A Catalytically Inactive Mutant (K223A) of C-DES with a Substrate (Cystine) Linked to the Co-Factor''' | ||
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[[Category: Mozzarelli, A.]] | [[Category: Mozzarelli, A.]] | ||
[[Category: Schiaretti, F.]] | [[Category: Schiaretti, F.]] | ||
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| - | [[Category: | + | [[Category: Inactive mutant]] |
| - | [[Category: | + | [[Category: Nifs-like]] |
| - | [[Category: | + | [[Category: Substrate complex]] |
| - | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 02:01:35 2008'' | |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | |
Revision as of 23:01, 2 May 2008
Structure of A Catalytically Inactive Mutant (K223A) of C-DES with a Substrate (Cystine) Linked to the Co-Factor
Overview
The cystine lyase (C-DES) of Synechocystis is a pyridoxal-5'-phosphate-dependent enzyme distantly related to the family of NifS-like proteins. The crystal structure of an N-terminal modified variant has recently been determined. Herein, the reactivity of this enzyme variant was investigated spectroscopically in solution and in the crystalline state to follow the course of the reaction and to determine the catalytic mechanism on a molecular level. Using the stopped-flow technique, the reaction with the preferred substrate cystine was found to follow biphasic kinetics leading to the formation of absorbing species at 338 and 470 nm, attributed to the external aldimine and the alpha-aminoacrylate; the reaction with cysteine also exhibited biphasic behavior but only the external aldimine accumulated. The same reaction intermediates were formed in crystals as seen by polarized absorption microspectrophotometry, thus indicating that C-DES is catalytically competent in the crystalline state. The three-dimensional structure of the catalytically inactive mutant C-DES(K223A) in the presence of cystine showed the formation of an external aldimine species, in which two alternate conformations of the substrate were observed. The combined results allow a catalytic mechanism to be proposed involving interactions between cystine and the active site residues Arg-360, Arg-369, and Trp-251*; these residues reorient during the beta-elimination reaction, leading to the formation of a hydrophobic pocket that stabilizes the enolimine tautomer of the aminoacrylate and the cysteine persulfide product.
About this Structure
1N31 is a Single protein structure of sequence from Synechocystis sp. pcc 6714. Full crystallographic information is available from OCA.
Reference
Snapshots of the cystine lyase C-DES during catalysis. Studies in solution and in the crystalline state., Kaiser JT, Bruno S, Clausen T, Huber R, Schiaretti F, Mozzarelli A, Kessler D, J Biol Chem. 2003 Jan 3;278(1):357-65. Epub 2002 Oct 16. PMID:12386155 Page seeded by OCA on Sat May 3 02:01:35 2008
