1n31: Difference between revisions
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|PDB= 1n31 |SIZE=350|CAPTION= <scene name='initialview01'>1n31</scene>, resolution 2.20Å | |PDB= 1n31 |SIZE=350|CAPTION= <scene name='initialview01'>1n31</scene>, resolution 2.20Å | ||
|SITE= | |SITE= | ||
|LIGAND= <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PLP:PYRIDOXAL-5'-PHOSPHATE'>PLP | |LIGAND= <scene name='pdbligand=CYS:CYSTEINE'>CYS</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PLP:PYRIDOXAL-5'-PHOSPHATE'>PLP</scene> | ||
|ACTIVITY= | |ACTIVITY= | ||
|GENE= C-DES ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id= | |GENE= C-DES ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1147 Synechocystis sp. PCC 6714]) | ||
|DOMAIN= | |||
|RELATEDENTRY=[[1elu|1ELU]], [[1elq|1ELQ]], [[1n2t|1N2T]] | |||
|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1n31 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1n31 OCA], [http://www.ebi.ac.uk/pdbsum/1n31 PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1n31 RCSB]</span> | |||
}} | }} | ||
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==About this Structure== | ==About this Structure== | ||
1N31 is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Synechocystis_sp. | 1N31 is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Synechocystis_sp._pcc_6714 Synechocystis sp. pcc 6714]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1N31 OCA]. | ||
==Reference== | ==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:[http://www.ncbi.nlm.nih.gov/pubmed/12386155 12386155] | 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:[http://www.ncbi.nlm.nih.gov/pubmed/12386155 12386155] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Synechocystis sp. pcc | [[Category: Synechocystis sp. pcc 6714]] | ||
[[Category: Bruno, S.]] | [[Category: Bruno, S.]] | ||
[[Category: Clausen, T.]] | [[Category: Clausen, T.]] | ||
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[[Category: Mozzarelli, A.]] | [[Category: Mozzarelli, A.]] | ||
[[Category: Schiaretti, F.]] | [[Category: Schiaretti, F.]] | ||
[[Category: fe-s cluster synthesis]] | [[Category: fe-s cluster synthesis]] | ||
[[Category: inactive mutant]] | [[Category: inactive mutant]] | ||
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[[Category: substrate complex]] | [[Category: substrate complex]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 22:24:38 2008'' | ||
Revision as of 22:24, 30 March 2008
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| , resolution 2.20Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | , , | ||||||
| Gene: | C-DES (Synechocystis sp. PCC 6714) | ||||||
| Related: | 1ELU, 1ELQ, 1N2T
| ||||||
| Resources: | FirstGlance, OCA, PDBsum, RCSB | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Structure of A Catalytically Inactive Mutant (K223A) of C-DES with a Substrate (Cystine) Linked to the Co-Factor
OverviewOverview
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 StructureAbout this Structure
1N31 is a Single protein structure of sequence from Synechocystis sp. pcc 6714. Full crystallographic information is available from OCA.
ReferenceReference
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
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