1q9i: Difference between revisions

From Proteopedia
Jump to navigationJump to search
← Older edit
OCA (talk | contribs)
Bots, Bureaucrats, journal, Administrators
3,332,843 edits
No edit summary
OCA (talk | contribs)
Bots, Bureaucrats, journal, Administrators
3,332,843 edits
No edit summary
 
(14 intermediate revisions by the same user not shown)
Line 1: Line 1:
[[Image:1q9i.jpg|left|200px]]
<!--
The line below this paragraph, containing "STRUCTURE_1q9i", creates the "Structure Box" on the page.
You may change the PDB parameter (which sets the PDB file loaded into the applet)
or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
or leave the SCENE parameter empty for the default display.
-->
{{STRUCTURE_1q9i|  PDB=1q9i  |  SCENE=  }}
'''The A251C:S430C double mutant of flavocytochrome c3 from Shewanella frigidimarina'''


==The A251C:S430C double mutant of flavocytochrome c3 from Shewanella frigidimarina==
<StructureSection load='1q9i' size='340' side='right'caption='[[1q9i]], [[Resolution|resolution]] 1.60&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[1q9i]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Shewanella_frigidimarina Shewanella frigidimarina]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1Q9I OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1Q9I FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.6&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=HEC:HEME+C'>HEC</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=TEO:MALATE+LIKE+INTERMEDIATE'>TEO</scene></td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1q9i FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1q9i OCA], [https://pdbe.org/1q9i PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1q9i RCSB], [https://www.ebi.ac.uk/pdbsum/1q9i PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1q9i ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/FCCA_SHEFN FCCA_SHEFN] Flavocytochrome that catalyzes the reduction of fumarate to succinate (PubMed:8093012). Is essential for fumarate respiration during anaerobic growth, acting as the terminal reductase (PubMed:9579067). Receives electrons from the membrane-bound tetraheme c-type cytochrome CymA (By similarity). Is essentially unidirectional, catalyzing only fumarate reduction (PubMed:8093012). Cannot reduce nitrite, dimethylsulphoxide, trimethylamine-N-oxide (TMAO) or sulfite (PubMed:8093012). In vitro, can use the artificial electron donor methyl viologen (PubMed:8093012).[UniProtKB:P83223]<ref>PMID:8093012</ref> <ref>PMID:9579067</ref>
== Evolutionary Conservation ==
[[Image:Consurf_key_small.gif|200px|right]]
Check<jmol>
  <jmolCheckbox>
    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/q9/1q9i_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.spt</scriptWhenUnchecked>
    <text>to colour the structure by Evolutionary Conservation</text>
  </jmolCheckbox>
</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1q9i ConSurf].
<div style="clear:both"></div>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
The crystal structures of various different members of the family of fumarate reductases and succinate dehydrogenases have allowed the identification of a mobile clamp (or capping) domain [e.g., Taylor, P., Pealing, S. L., Reid, G. A., Chapman, S. K., and Walkinshaw, M. D. (1999) Nat. Struct. Biol. 6, 1108-1112], which has been proposed to be involved in regulating accessibility of the active site to substrate. To investigate this, we have constructed the A251C:S430C double mutant form of the soluble flavocytochrome c(3) fumarate reductase from Shewanella frigidimarina, to introduce an interdomain disulfide bond between the FAD-binding and clamp domains of the enzyme, thus restricting relative mobility between the two. Here, we describe the kinetic and crystallographic analysis of this double mutant enzyme. The 1.6 A resolution crystal structure of the A251C:S430C enzyme under oxidizing conditions reveals the formation of a disulfide bond, while Ellman analysis confirms its presence in the enzyme in solution. Kinetic analyses with the enzyme in both the nonbridged (free thiol) and the disulfide-bridged states indicate a slight decrease in the rate of fumarate reduction when the disulfide bridge is present, while solvent-kinetic-isotope studies indicate that in both wild-type and mutant enzymes the reaction is rate limited by proton and/or hydride transfer during catalysis. The limited effects of the inhibition of clamp domain mobility upon the catalytic reaction would indicate that such mobility is not essential for the regulation of substrate access or product release.


==Overview==
Probing domain mobility in a flavocytochrome.,Rothery EL, Mowat CG, Miles CS, Mott S, Walkinshaw MD, Reid GA, Chapman SK Biochemistry. 2004 May 4;43(17):4983-9. PMID:15109257<ref>PMID:15109257</ref>
The crystal structures of various different members of the family of fumarate reductases and succinate dehydrogenases have allowed the identification of a mobile clamp (or capping) domain [e.g., Taylor, P., Pealing, S. L., Reid, G. A., Chapman, S. K., and Walkinshaw, M. D. (1999) Nat. Struct. Biol. 6, 1108-1112], which has been proposed to be involved in regulating accessibility of the active site to substrate. To investigate this, we have constructed the A251C:S430C double mutant form of the soluble flavocytochrome c(3) fumarate reductase from Shewanella frigidimarina, to introduce an interdomain disulfide bond between the FAD-binding and clamp domains of the enzyme, thus restricting relative mobility between the two. Here, we describe the kinetic and crystallographic analysis of this double mutant enzyme. The 1.6 A resolution crystal structure of the A251C:S430C enzyme under oxidizing conditions reveals the formation of a disulfide bond, while Ellman analysis confirms its presence in the enzyme in solution. Kinetic analyses with the enzyme in both the nonbridged (free thiol) and the disulfide-bridged states indicate a slight decrease in the rate of fumarate reduction when the disulfide bridge is present, while solvent-kinetic-isotope studies indicate that in both wild-type and mutant enzymes the reaction is rate limited by proton and/or hydride transfer during catalysis. The limited effects of the inhibition of clamp domain mobility upon the catalytic reaction would indicate that such mobility is not essential for the regulation of substrate access or product release.


==About this Structure==
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
1Q9I is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Shewanella_frigidimarina Shewanella frigidimarina]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1Q9I OCA].
</div>
<div class="pdbe-citations 1q9i" style="background-color:#fffaf0;"></div>


==Reference==
==See Also==
Probing domain mobility in a flavocytochrome., Rothery EL, Mowat CG, Miles CS, Mott S, Walkinshaw MD, Reid GA, Chapman SK, Biochemistry. 2004 May 4;43(17):4983-9. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15109257 15109257]
*[[Flavocytochrome 3D structures|Flavocytochrome 3D structures]]
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Large Structures]]
[[Category: Shewanella frigidimarina]]
[[Category: Shewanella frigidimarina]]
[[Category: Single protein]]
[[Category: Chapman SK]]
[[Category: Succinate dehydrogenase]]
[[Category: Miles CS]]
[[Category: Chapman, S K.]]
[[Category: Mowat CG]]
[[Category: Miles, C S.]]
[[Category: Reid GA]]
[[Category: Mowat, C G.]]
[[Category: Rothery EL]]
[[Category: Reid, G A.]]
[[Category: Walkinshaw MD]]
[[Category: Rothery, E L.]]
[[Category: Walkinshaw, M D.]]
[[Category: Disulfide]]
[[Category: Flavocytochrome]]
[[Category: Fumarate reductase]]
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May  3 06:02:09 2008''
Retrieved from "https://proteopedia.org/1q9i"